Practical Chemistry Techniques for Success

Practical Skills in
Chemistry
Pearson
Education
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First published 2002
© Pearson Education Limited 2002
The rights of John R. Dean, Alan M. Jones, David Holmes,

Rob Reed, Jonathan Weyers and Allan Jones to be identified
as authors of this work have been asserted by them in
accordance with the Copyright, Design and Patents Act 1988.
All rights reserved; no part of this publication may be

reproduced, stored in any retrieval system, or transmitted
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Copyright Licensing Agency Ltd, 90 Tottenham Court Road,
London wn 4LP.
ISBN 978-0-13-028002-2
British Library Cataloguing-in-Publication Data

A catalogue record for this book is available from the British Library
10 9 8 7 6
08 07
Typeset in Times Roman by 32

Printed and bound in Great Britain by Ashford Colour Press
Gosport, Hants.
Contents
Boxes vii
Preface IX
Acknowledgements X
Abbreviations xiii
Fundamental laboratory techniques

1 Basic principles 3
2 Health and safety 6
3 Working with liquids 9
4 Basic laboratory procedures I 15
5 Basic laboratory procedures 11 26
6 Principles of solution chemistry 45
7 pH and buffer solutions 56
Resources for fundamental laboratory techniques 62
The investigative approach

8 Making and recording measurements 65
9 SI units and their use 70
10 Scientific method and design of experiments 75
11 Project work 80
Resources for the investigative approach 83
Laboratory techniques
12 Melting points 87
13 Recrystallization 92
14 Solvent extraction 102
15 Distillation 107
16 Reflux 116
17 Evaporation 121
18 Inert atmosphere methods 125
Resources for laboratory techniques 131
Classical techniques
19 Qualitative techniques for inorganic analysis 135
20 Gravimetry 139
21 Procedures in volumetric analysis 141
22 Acid-base titrations 148
23 Complexometric titrations 151
24 Redox titrations 155
25 Precipitation titrations 157
Resources for classical techniques 160
Instrumental techniques
26 Basic spectroscopy 163
27 Atomic spectroscopy 170
28 Infrared spectroscopy 180
29 Nuclear magnetic resonance spectrometry 190
30 Mass spectrometry 200
31 Chromatography ~ introduction 205
32 Gas and liquid chromatography 211
v
Contents
33 Electrophoresis 225
34 Electroanalytical techniques 229
35 Radioactive isotopes and their uses 235
36 Thermal analysis 243
Resources for instrumental techniques 246
Analysis and presentation of data

37 U sing graphs 251
38 Presenting data in tables 256
39 Hints for solving numerical problems 259
40 Descriptive statistics 264
41 Choosing and using statistical tests 271
42 Drawing chemical structures 280
43 Chemometrics 285
44 Computational chemistry 291
Resources for analysis and presentation of data 294
Information technology and library resources

45 The Internet and World Wide Web 299
46 Internet resources for chemistry 302
47 Using spread sheets 307
48 Word processors, databases and other packages 311
49 Finding and citing published information 317
Communicating information
50 General aspects of scientific writing 325
51 Writing essays 330
52 Reporting practical and project work 332
53 Writing literature surveys and reviews 339
54 Organizing a poster display 341
55 Giving an oral presentation 344
56 Examinations 348
Resources for communicating information 354
References 355
Index 357
vi
Boxes
4.1 How to make up an aqueous solution of known concentration from a

solid chemical 17
4.2 How to make up an aqueous solution of known concentration from a
solid chemical for use in quantitative analysis 19
4.3 How to make up a linear dilution series for use in quantitative analysis 21
4.4 How to weigh out a sample of a solid for use in quantitative analysis 24
5.1 How to flute a filter paper for gravity filtration 28
5.2 Isolation of a solid by suction filtration 30
5.3 How to dry a solution over magnesium sulphate 41
6.1 Useful procedures for calculations involving molar concentrations 46
6.2 How to convert ppm into mass of chemical required 47
6.3 An example of complex formation 53
6.4 The use of oxidation numbers to identify redox systems 54
6.5 How to balance redox equations from partial ionic equations using the
ion-electron method 55
7.1 Using a glass pH electrode and meter to measure the pH of a solution 59
9.1 Conversion factors between some redundant units and the SI 72
9.2 How to interconvert SI units 73
10.1 Checklist for designing and executing an experiment 77
13.1 How to carry out a solvent selection for recrystallization of an unknown
compound 94
13.2 How to carry out a mixed-solvent selection for recrystallization of an
unknown compound 95
13.3 How to carry out a single-solvent recrystallization 97
13.4 How to carry out a mixed-solvent recrystallization 99
14.1 How to separate a carboxylic acid and a hydrocarbon using solvent
extraction 105
14.2 How to set up a Soxh1et extraction system 106
15.1 How to assemble the apparatus for a simple distillation 109
15.2 How to carry out a simple distillation 109
15.3 How to carry out a fractional distillation 110
15.4 How to carry out a reduced-pressure distillation using a water pump 113
15.5 How to carry out a steam distillation 114
16.1 How to set up a simple reflux apparatus 117
16.2 How to set up the apparatus for reflux with mechanical stirring 118
17.1 How to use a rotary film evaporator 122
18.1 How to transfer an air-sensitive reagent using a syringe 130
19.1 How to use a low-speed bench centrifuge 137
20.1 How to carry out gravimetric analysis 139
21.1 Types of calculations used in volumetric analysis - titrations 142
26.1 Using a UV /visible spectrophotometer 166
26.2 U sing a flame photometer 169
27.1 How to prepare a 1000 J.lgmL-1 stock solution of a metal ion from a
metal salt 170
27.2 How to prepare a set of five calibration solutions in the concentration
range Os-ltlzzg mL-1(mgL-1) 171
27.3 How to analyse a sample using the method of standard additions in
FAAS 172
27.4 Sample size and certified reference materials 173
vii
Boxes
27.5 Analysis of a sample: dilution factor 173

27.6 How to operate a flame atomic absorption spectrometer 177
27.7 How to acid-digest a sample using a hot plate 179
28.1 How to run an infrared spectrum of a liquid or solid film, mull or
KBr disk 183
28.2 How to prepare liquid and solid films and mulls 185
28.3 How to prepare a KBr disk 188
28.4 How to interpret an IR spectrum 189
29.1 How to prepare a sample for NMR spectroscopy 193
30.1 How to identify the number of bromine or chlorine atoms in a
molecule from the molecular ion 202
30.2 Idealized fragmentation processes for the molecular ion (M+) 203
32.1 How to prepare a set of five calibration solutions in the concentration
range O-IQ IlgmL -I (mgL -I) 212
32.2 How to make micro pipettes for TLC 217
32.3 How to run a thin-layer chromatogram 218
35.1 How to determine the specific activity of an experimental solution 238
35.2 Tips for preparing samples for liquid scintillation counting 239
36.1 How to interpret a thermal analysis trace 244
37.1 Checklist for the stages in drawing a graph 253
38.1 Checklist for preparing a table 257
39.1 Example of using the rules of Table 39.2 261
40.1 Calculation of descriptive statistics for a sample of grouped data 265
40.2 Three examples where simple arithmetic means are inappropriate 266
41.1 How to carry out at-test 276
41.2 Worked example of at-test 277
43.1 Example of a two-level factorial design 287
43.2 Example of principal component analysis 290
45.1 Important guidelines for using computers and networks 301
50.1 How to achieve a clear, readable style 327
50.2 Improve your writing ability by consulting a personal reference library 328
52.1 The structure of reports of experimental work 333
52.2 Writing experimental procedures 335
52.3 Steps in producing a scientific paper 337
56.1 Writing under exam conditions 352
viii
Preface
Practical skills form the cornerstone of chemistry. However, the diversity of

skills required in the laboratory means that a student's experience may be
limited. While some techniques do require specific skills, many of them are
transferable, generic skills that are required throughout the subject area.
Limited time constraints of the modern curriculum often preclude or
minimize laboratory time. It is the aim of this book to provide general
guidance for use in and outside practical sessions and also to cover a range of
techniques from the basic to the more advanced.
The book is primarily aimed at the undergraduate level (principally first
and second year), although some information is relevant to final year
undergraduates and postgraduate courses. The diversity of the book means
that recipe-like solutions are not provided for every potential problem, but
guidance on a range of topics is provided.
As the book focuses on practical aspects it is important not to apportion
'labels' to the areas covered. However, for the traditionalist the areas covered
are primarily focused on synthetic, physical and analytical aspects of
chemistry.
It is intended that the book will be useful in the laboratory, during
practical classes, and during project work. It is not intended to replace recipe-
like laboratory scripts but to enable students to become familiar with the
practical aspects of the subject both at the time of performing the experiment
and during the writing-up process. In addition, lecturers should find that the
text provides an effective means of supplementing the information given in
practical classes, where constraints on time and resources can lead to the
under-performance of students.
We have selected material for inclusion in Practical Skills in Chemistry
based on our own teaching experience, highlighting those areas where our
students have needed further guidance. As a result of our comprehensive
cover of practical skills, some techniques such as microscale methods and
specialized vacuum techniques have been omitted, but specific references are
provided. Instead, we have attempted to provide sufficient detail so that
students will have the skills to carry out experiments successfully and not
produce poor data as a result of poor technique.
Most students will have access to specialist textbooks giving in-depth
coverage of the theoretical principles and knowledge covered in lectures. This
book aims to supplement - rather than replace - such textbooks, and covers
the skills required in laboratory classes, together with practical advice, tips,
hints, worked examples, definitions, key points, 'how to' boxes and checklists
which are useful in the field of chemistry. The text provides an outline on the
underlying theoretical principles where necessary, but emphasis throughout is
on the practical applications of this information.
To students who buy this book, we hope that you will find it useful in the
laboratory, during your practical classes and in your project work - this is
not a book to be left on the bookshelf. Lecturers should find that the text
provides an effective means of supplementing the information given in
practical classes.
We would like to acknowledge the support of our families and the help
provided by colleagues who read early drafts of the material. In particular,
special thanks are due to Gary Askwith, Dave Bannister (Manchester
ix
Preface
Metropolitan University), Jon Bookham, Susan Carlile, Jim Creighton, Sarah

Cresswell, Martin Davies, Les Dix, Jackie Eager, Derek Holmes, Ed Ludkin,
Dave Osborne, Justin Perry, Jane Shaw, Tony Simpson, Dave Wealleans and
Ian Winship. Despite this help, the responsibility for any errors rests with us
and we would be grateful if readers could alert us to any errors or problems
so that we can make amends as soon as possible. Please write to us at the
University of Northumbria at Newcastle, or bye-mail at the following
addresses:
[email protected] (JRD);
[email protected] (Alan M. Jones);
[email protected] (RHR);
[email protected] (DH);
[email protected] (JDBW);
[email protected] (Allan Jones).
Acknowledgements
We are grateful to the following for permISSIOn to reproduce copyright

material:
Text extracts from the following databases: Analytical Abstracts, Chemical

Business NewsBase, Chemical Safety NewsBase, Mass Spectrometry Bulletin,
Chemical Safety Data Sheets and Chromatography Abstracts in Chapter 36,
'Internet resources for chemistry', reproduced by kind permission of The
Royal Society of Chemistry.
Whilst every effort has been made to trace the owners of copyright material,
in a few cases this has proved impossible and we take this opportunity to
offer our apologies to any copyright holders whose rights we may have
unwittingly infringed.
x
For the student
This text is designed to help you in your Chemistry studies, before and
during laboratory classes, in project work, in tutorials and even in
examinations.
Chapters 1-36 cover general and specific skills for laboratory work.
These are based on the authors' experience of questions that students often
ask and difficulties they have in performing laboratory work. They include
tips, hints, worked examples, definitions and "how to" boxes that set out
procedures for you step by step.
Chapters 37-44 explain data analysis and presentation.
This will be an important element of your course and you will find that this
section guides you through the key skills that you will need to develop, from
presenting graphs and tables to drawing chemical structures and using
statistical tests.
Chapters 45-49 cover information technology and library resources.
These chapters will help you get the most from chemistry resources available
on the World Wide Web.
Chapters 50-56 deal with the vital subject of communicating information.
Designed to help you report practical and project work and give oral
presentations effectively, and to succeed in examinations.
We hope you will find this book a helpful guide throughout your course, and
beyond.
xi
Abbreviations
Ar relative atomic mass

AAS atomic absorption spectrometry
AC affinity chromatography
ACS American Chemical Society
AES atomic emission spectrometry
ANOVA analysis of variance
AO atomic orbital
ATP adenosine triphosphate
BIDS Bath Information and Data Services
b.pt. boiling point
CCD central composit design
CCP cubic close packed
CE capillary electrophoresis
CEC capillary electrochromatography
CGE capillary gel electrophoresis
COSHH Control of Substances Hazardous to Health
CoY coefficient of variance
CRm certified reference material
CZE capillary zone electrophoresis
DAD diode array detection
DCM dichloromethane
DNA deoxyribonucleic acid
DSC differential scanning colorimetry
DTA differential thermal analysis
ECD electron capture detector
EDTA ethylenediaminetetraacetic acid
El electron impact ionization
EIE easily ionizable element
EMR electromagnetic radiation
en ethylenediamine
EOF electro-osmotic flow
F Faraday constant
FAAS flame atomic absorption spectrometer
FID flame ionization detector
FT Fourier transformation
FT-IR Fourier transform - infrared (spectroscopy)
GC gas chromatography
GFC gel filtration chromatography
GPC gel permeation chromatography
h Planck constant
HASAW Health and Safety at Work
HCB hexachloro-l,3-butadiene
HCL hollow-cathode lamp
HCP hexagonal close packed
HIC hydrophobic interaction chromatography
HPLC high-performance liquid chromatography
ICP inductively coupled plasma
IEC ion-exchange chromatography
IEF isoelectric focusing
xiii
Abbreviations
IR infrared (radiation)
ISE ion selective electrode
IUPAC International Union of Pure and Applied Chemistry
s; acid dissociation constant
s, solubility product
Kw ion product of water
KHP potassium hydrogen phthalate
LGC Laboratory of the Government Chemist
u, relative molecular mass
MDL minimum detectable level
MEKC micellar electrokinetic chromatography
MEL maximum exposure limit
MO molecular orbital
m.pt. melting point
MS mass spectrometry
NH null hypothesis
NIST National Institute of Standards and Technology
NMR nuclear magnetic resonance
NP-HPLC normal phase high-performance liquid chromatography
ODA octadecy lsilane
OEL occupational exposure standard
PCA principal component analysis
PFA perfluoroalkoxyvin ylether
PLOT porous layer open tubular (column)
PMT photomultiplier tube
PTFE polytetrafluoroethylene
R universal gas constant
Rr relative frontal mobility
RA relative abundance
RNA ribonucleic acid
RP-HPLC reverse phase high-performance liquid chromatography
rpm revolutions per minute
RSC Royal Society of Chemistry
SAX strong anion exchange
SCOT support-coated open tubular (capillary column)
SCX strong cation exchange
SDS sodium dodecyl sulphate
SE standard error (of the sample mean)
SEM scanning electron microscopy
SI Systeme Internationale d'Unites
STP standard temperature and pressure
TCA trichloracetic acid
TCD thermal conductivity detector
TG thermogravimetry
TLC thin-layer chromatography
TMS tetramethylsilane
TRIS tris(hydroxymethyl)aminomethane or
2-amino- 2-hydroxymethy 1-1,3-propanediol
URL uniform resource locator
USEPA United States Environmental Protection Agency
UV ultraviolet
WCOT wall-coated open tubular (column)
WWW World Wide Web
xiv
-------0 Basic principles
All knowledge and theory in science has originated from practical

observation and experimentation: this is equally true for disciplines as
diverse as analysis and synthesis. Laboratory work is an essential part of
all chemistry courses and often accounts for a significant proportion of
the assessment marks. This book aims to provide an easy-to-use reference
source dealing with basic practical techniques and information. The skills
developed in practical classes will continue to be useful throughout your
course and beyond, some within science and others in any career you
choose.
Being prepared
~ You will get the most out of laboratory work if

you prepare well. Do not go into a practical session assuming that
everything will be provided, without any input on your part.
The main points to remember are:
• Read any handouts in advance: make sure you understand the purpose
of the practical and the particular skills involved. Does the practical
relate to, or expand upon, a current topic in your lectures? Is there any
additional preparatory reading that will help?
• Take along appropriate textbooks, to explain aspects in the practical.
• Consider what safety hazards might be involved, and any precautions
you might need to take, before you begin (p. 7).
• Listen carefully to any instructions and note any important points:
adjust your schedule/handout, as necessary.
• During the practical session, organize your bench space - make sure
your lab book is adjacent to, but not within, your working area. You
will often find it easiest to keep clean items of glassware etc. on one side
of your working space, with used equipment on the other side.
• All chemical waste (solid or liquid) should be disposed of in the
appropriate containers provided (consult the demonstrator or lecturer-
in-charge).
• Write up your work as soon as possible, and submit it on time, or you
may lose marks.
• Catch up on any work you have missed as soon as possible - preferably
before the next practical session.
Basic requirements
Recording practical results
An A4 loose-leaf ring binder offers flexibility, since you can insert
laboratory handouts, and lined and graph paper, at appropriate points. The
danger of losing one or more pages from a loose-leaf system is the main
drawback. Bound books avoid this problem, although those containing
alternating lined/graph or lined/blank pages tend to be wasteful - it is often
better to paste sheets of graph paper into a bound book, as required.
Fundamental laboratory techniques 3

Basic principles
All experimental observations and data should be recorded in a notebook

in ink at the time they are made because it is easy to forget when you are
busy.
A good-quality HB pencil or propelling pencil is recommended for
making diagrams etc. as mistakes are easily corrected with a vinyl eraser.
Buy a black, spirit-based (permanent) marker to label experimental
glassware, sample tubes, etc. Fibre-tipped fine line drawing/lettering pens
are useful for preparing final versions of graphs and diagrams for
assessment purposes. Use a clear ruler (with an undamaged edge) for graph
drawing, so that you can see data points/information below the ruler as
you draw.
Calculators
These range from basic machines with no pre-programmed functions and
only one memory, to sophisticated programmable minicomputers with
many memories. The following may be helpful when using a calculator:
• Power sources. Choose a battery-powered machine, rather than a mains-
operated or solar-powered type. You will need one with basic mathe-
matical/scientific operations including powers, logarithms (p. 262), roots
and parentheses (brackets), together with statistical functions such as
sample means and standard deviations (Chapter 40).
• Mode of operation. Calculators fall into two distinct groups. The older
system used by, for example, Hewlett Packard calculators is known as
the reverse Polish notation: to calculate the sum of two numbers, the
sequence is 2 [enter] 4 + and the answer 6 is displayed. The more usual
method of calculating this equation is as 2 + 4 =, which is the system
used by the majority of modern calculators. Most newcomers find the
latter approach to be more straightforward. Spend some time finding
out how a calculator operates, e.g. does it have true algebraic logic (..j
then number, rather than number then ..j)? How does it deal with
scientific notation (p. 262)?
• Display. Some calculators will display an entire mathematical operation
(e.g. '2 + 4 = 6'), while others simply display the last number/operation.
The former type may offer advantages in tracing errors.
• Complexity. In the early stages, it is usually better to avoid the more
complex machines, full of impressive-looking, but often unused pre-
programmed functions - go for more memory, parentheses, or statistical
functions rather than engineering or mathematical constants. Program-
mable calculators may be worth considering for more advanced studies.
However, it is important to note that such calculators are often
unacceptable for exams.
Presenting more advanced practical work

In some practical reports and in project work, you may need to use more
sophisticated presentation equipment. Word processing may be essential
and computer-based graphics packages can be useful. Choose easily-read
fonts such as Arial or Times New Roman for project work and posters and
consider the layout and content carefully (p. 341). Alternatively, you could
use fine line drawing pens plus dry-transfer lettering and symbols, such as
those made by Letraset'v, although this approach is usually more time
consuming and less flexible than computer-based systems.
4 Fundamental laboratory techniques

Basic principles
To prepare overhead transparencies for oral presentations, you can use

spirit-based markers and acetate sheets. An alternative approach is to print
directly from a computer-based package, using a laser printer and special
acetates, or directly to 35 mm slides. You can also photocopy on to special
acetates. The use of Microsoft Powerf'oint'P as a presentation package has
become more important in recent years. It is not uncommon to find a
computer and presenter available for student use. Advice on content and
presentation is given on p. 344.

------8 Health and safety
Health and safety law requires institutions to provide a working environ-

ment that is safe and without risk to health. Where appropriate, training
and information on safe working practices must be provided. Students and
staff must take reasonable care to ensure the health and safety of
themselves and of others, and must not misuse any safety equipment.
~ All practical work must be carried out with safety in

mind, to minimize the risk of harm to yourself and to others - safety is
everyone's responsibility by law.
Risk assessment
The most widespread approach to safe working practice involves the use of
risk assessment, which aims to establish:
1. The intrinsic chemical and physical hazards, together with any maximum
exposure limits (MELs) or occupational exposure standards (OESs),
where appropriate. All chemical manufacturers provide data sheets
listing the hazards associated with particular chemical compounds.
2. The risks involved, by taking into account the amount of substance to be
used, the way in which it will be used and the possible routes of entry
into the body (Fig. 2.1). In this regard, it is important to distinguish
between the intrinsic hazards of a particular substance and the risks
involved in its use in a particular exercise.
3. The persons at risk and the ways in which they might be exposed to
hazardous substances, including accidental exposure (spillage).
4. The steps required to prevent or control exposure. Ideally, a non-
hazardous or less hazardous alternative should be used. If this is not
feasible, adequate control measures must be used, e.g. a fume cupboard
or other containment system. Personal protective equipment (e.g. lab
coats, safety glasses) must continue to be used in addition to such
containment measures. A safe means of disposal will be required.
The outcome of the risk assessment process must be recorded and
appropriate safety information must be passed on to those at risk. For most
practical classes, risk assessments will have been carried out in advance by the
inoculation person in charge and the information necessary to minimize the risks to
or
absorption
students may be given in the practical schedule. You will be asked to carry out
risk assessments to familiarize yourself with the process and sources of
information. Make sure you know how your department provides such
information and that you have read the appropriate material before you begin
your practical work. You should also pay close attention to the person in
charge at the beginning of the practical session, as they may emphasize the
major hazards and risks. In project work, you will need to be involved in the
risk assessment process along with your supervisor, before you carry out any
laboratory work. Any new materials synthesized during the project should be
treated with the utmost respect. An example of a risk assessment is shown in
Fig. 2.2.
Fig. 2.1 Major routes of entry of harmful In addition to specific risk assessments, most institutions will have a safety
substances into the body. handbook, giving general details of safe working practices, together with the

Health and safety
LABORATORY RISK ASSESSMENT
This sheet must be completed before commencing each experiment
I Name:
I Experiment: Synthesis of N-phenylethanamide Date:
t. Hazard and Risk Codes Enter below the correct Hazard Code and Risk Code for each reagent, solvent,
product and by product (letter and number in the relevant boxes)
REAGENT AND HAZARD (H) and RISK (R) CODES

INSTRUMENTS
Others
(insert)
A C T F W M
H R H R H R H R H R H R H R H R
Aminobenzene (aniline) 2 2 2 2 2 2 2 1 1 1 1 1
Ethanoic anhydride 2 2 1 1 2 2 2 1 2 1 1 1
N-phenylethanamide 1 1 1 1 2 1 1 1 1 1 1 1
Ethanoic acid (dilute aqueous solution) 1 1 1 1 1 1 1 1 1 1 1 1
A = Corrosive/irritant M = Microbiological hazard 1 =low/nohazardJrisk

C = Carcinogenic R = Radioactive 2=Moderatehazardlrisk
T = Toxic I = Instrument hazard 3 = High hazard/risk
F=Flammable X = Explosive 4 = Special high category
W = Violent reaction with water/acid/bases 0= Oxidizing agents
2. Precautions: containment and protection
Stage of Experiment Containment Personal Additional Information

(including disposal) Code Protection
Weighing out chemicals 3 Gloves
Containment code 1 = Open Lab 2 = Restricted lab (no naked flames) 3 = Fume Cupboard 4 = Glove Box/Safety Cabinet
Personal Protection Insert all requirements additional to coat and glasses (e.g. gloves, visor, safety screen, etc.I
Additional Information For example, disposaVspecffied waste container, sink and water, etc.
3. Signatures
Student: Supervisor: Date:
.
----
Fig.2.2 An example of a laboratory hazard assessment form.
names and telephone numbers of safety personnel, first aiders, hospitals, etc.
Make sure you read this and abide by any instructions.
Basic rules for laboratory work

• Wear appropriate protective clothing at all times - a clean lab coat
(buttoned up), eye protection, appropriate footwear and ensure your hair
does not constitute a hazard.
• Never smoke, eat or drink in any laboratory, because of the risks of
contamination by inhalation or ingestion (Fig. 2.1).
• Never work alone in a laboratory.
• Make sure that you know what to do in case of fire, including exit routes,
how to raise the alarm, and where to gather on leaving the building.
Remember that the most important consideration is human safety: do not
attempt to fight a fire unless it is safe to do so.

Health and safety
• All laboratories display notices telling you where to find the first aid kit
and who to contact in case of accident/emergency. Report all accidents,
even those appearing insignificant - your department will have a
reporting procedure to comply with safety legislation.
• Know the warning symbols for specific chemical hazards (Fig. 2.3).
• Never touch chemicals unless they are known to have minimal hazard:
use a spatula to transfer and manipulate solids, and pipettes for liquids -
see p. 9.
• Never mouth pipette any liquid. Use a pipette filler (see p. 10).
• Take care when handling glassware - see p. 13 for details.
• Use a fume cupboard for hazardous chemicals. Make sure that it is
working and then open the front only as far as necessary: many fume
cupboards are marked with a maximum opening.
• Always use the minimum quantity of any hazardous materials.
• Work in a logical, tidy manner and minimize risks by thinking ahead.
• Alway clear up spillages, especially around balances, infrared sample
preparation areas, etc., for the next worker.
• Always clear up at the end of each session. This is an important aspect of
safety, encouraging a responsible attitude towards laboratory work.
Fig.2.3 Warning labels for specific chemical

hazards.

------~8 Working with liquids
Measuring and dispensing liquids

The equipment you should choose to measure out liquids depends upon the
volume to be dispensed, the accuracy required and the number of times the
job must be repeated (Table 3.1).
Table 3. 1 Criteria for choosing a method for measuring out a liquid
Pasteur pipette 1-5 mL Low Convenient

Conical flask/beaker 25-5000 mL Very low Convenient
Measuring cylinder 5-2000mL Medium Convenient
Volumetric flask 5-2000 mL High Convenient
Burette 1-100mL High Convenient
Glass pipette 1-100mL High Convenient
Mechanical pipettor 5-1000,uL High* Convenient
Syringe 0.5-20,uL Medium** Convenient
Microsyringe 0.5-50,uL High Convenient
Weighing Any (depends Very high Inconvenient
on accuracy
of balance
*If calibrated correctly and used properly (see p. 10).

**Accuracy depends on width of barrel: large volumes less accurate.
Conical flasks, beakers, measuring cylinders and volumetric flasks

measure the volume of liquid contained in them, while burettes, pipettes,
pipettors, syringes and micro syringes mostly measure the volume delivered
from them: think about the requirements of the experiment.
Certain liquids may cause problems:
• High-viscosity liquids are difficult to dispense: allow time for the liquid to
transfer.
• Organic solvents may evaporate rapidly, making measurements inac-
curate: work quickly; seal containers quickly.
thumb and index • Solutions prone to frothing (e.g. surfactant solutions) are difficult to
finger provide
pressure on bulb measure and dispense: avoid forming bubbles; do not transfer quickly.
Pasteur pipettes
Hold correctly during use (Fig. 3.1) - keep the pipette vertical, with the
middle finger gripping the barrel to support the pipette while the thumb and
index finger provide controlled pressure on the bulb, and squeeze gently to
provide individual drops.
To prevent liquid being sucked into the bulb and hence cross-contamination:
• Ensure that the capacity of the bulb does not exceed that of the barrel.
• Do not remove the tip of the pipette from the liquid while drawing up the
liquid; the inrush of air may splash the liquid into the bulb. This is
particularly true when you lose patience trying to draw up viscous liquids.
• Do not lie the pipette on its side during use.
Conversely, if volatile liquids such as dichloromethane (DCM), ethanol,
Fig. 3.1 How to hold a Pasteur pipette. propanone (acetone) or diethylether (ether), for example, are to be

Working with liquids
dispensed, the warmth of the glass pipette will cause the liquid to squirt
from the pipette without any pressure on the bulb. To prevent this, suck up
the liquid several times into the pipette so as to cool the glass and then
dispense as normal.
Conical flasks and beakers

These have approximate graduations and should only be used for
10 measuring volumes of solutions/liquids where accuracy is unimportant.
Measuring cylinders and volumetric flasks

These must be used on a level surface (the laboratory bench) so that the
4
25 scale is horizontal; you should first fill with solution until just below the
desired mark, then fill slowly (e.g. using a Pasteur pipette) until the bottom
of the meniscus is level with the mark. Remember to allow time for the
solution to run down the walls of the vessel and to bend down so that your
eyes are level with the graduation mark(s) and the meniscus.
Burettes
These must be mounted vertically in a clamp - don't over-tighten the clamp
10 (see p. 26) - or in a burette holder, on a stand. First ensure that the tap is
closed and, using a funnel, add a little of the solution to be dispensed, rinse
the burette and discard the washings through the tap: this is vital in
titrations where a little water in the burette will alter the concentration of
la) (bl (c) the solution. Refill the burette with solution, open the tap and allow the
liquid to fill the barrel below the tap, then take a meniscus reading, noting
Fig. 3.2 Glass pipettes: (a) graduated pipette,
reading from zero to shoulder; (b) graduated
the value in your notebook. Dispense the solution via the tap and measure
pipette, reading from maximum to tip, by the new meniscus reading. The volume dispensed is the difference between
gravity; (c) bulb (volumetric) pipette, showing the two readings.
volume on bulb.
Pipettes
There are various designs, including graduated and bulb (volumetric)
pipettes (Fig. 3.2). Take care to look at the volume scale before use: some
graduated pipettes empty from full volume to zero, others from zero to full
volume; some scales refer to the shoulder of the tip, others to the tip by
gravity. Never blowout volumetric (bulb) pipettes, just touch the tip
against the inside wall of the vessel.
Rinse out pipettes with a little of the solution to be delivered before
commencing the accurate measurement. To prevent cross-contamination,
never draw the solution into the pipette filler.
~ For safety reasons, it is no longer permissible to

mouth pipette - various aids (pipette fillers) are available, such as the
rubber-bulb and Pi-Pump® (Fig. 3.3).
Pipettors (autopipettors)
There are two basic types:
1. Air displacement pipettors. For routine work with dilute aqueous sol-
utions. One of the most widely used is the Gilson Pipetrnan'f (Fig. 3.4).
2. Positive displacement pipettors. For non-standard applications, including
dispensing viscous, dense or volatile liquids where an air displacement
pipettor might create aerosols leading to errors.

Air displacement and positive displacement pipettors may be:

evacuate
• Fixed volume: capable of delivering a single factory-set volume.
bulb
• Adjustable: where the volume delivered is determined by the operator
across a particular range of values.
• Pre-set: movable between a limited number of values.
• Multichannel: able to deliver several replicate volumes at the same time.
Whichever type of these routine but expensive devices you use, you must
ensure that you understand the operating principles of the volume scale and
the method for changing the volume delivered - some pipettors are easily
misread.
empty A pipettor must be fitted with the correct disposable tip before use and
pipette
each manufacturer produces different tips to fit particular models. Special-
ized tips are available for particular applications.
(a) (b) If you accidentally draw liquid into the barrel, seek assistance from your
demonstrator/supervisor since the barrel will need to be cleaned before
Fig. 3.3 Pipette fillers: (a) rubber-bulb type; (b)
further use (to prevent cross-contamination) and unskilled dismantling of
Pi-Pump@.
the device will cause irreparable damage.
Syringes
Syringes should be used by placing the tip of the needle into the solution
and slowly drawing the plunger up to the required point on the scale. Check
the barrel to make sure no air bubbles have been drawn up, and expel the
solution slowly, touching the needle tip on the side of the vessel to remove
any adhering solution. If there is air in the barrel, fill past the mark, invert
the syringe and push the plunger to the mark so that the air and a little of
push button the solution are expelled into a waste collection vessel. Then dispense the
plunger solution. The use of syringes for dispensing air-sensitive reagents is
tip ejector button
described in Chapter 18.
adjustment ring
Microsyringes should always be cleaned before and after use by
repeatedly drawing up and expelling pure solvent. The dead space in the
volume scale
(volumeter needle can occupy up to 4% of the nominal syringe volume. Some micro-
syringes have a fine wire attached to the plunger, which fills the dead space.
Never pull the plunger out of the barrel.
barrel Balances
These can be used to weigh accurately (p. 22) how much liquid you have
dispensed. Convert mass to volume using the equation:
Mass/density = volume
tip ejector
e.g. a liquid (9.0g) of density (l.2gmL~I) = 7.5mL. Densities of common
solvents and common chemicals can be found in Lide (2000). You will also
disposable tip need to know the liquid's temperature, since density is temperature de-
pendent.
Fig. 3.4 A pipettor - the Gilson Pipetman'P.

Holding and storing liquids
Test tubes
Both 'normal' and the much smaller 'semi-micro' test tubes are used for
small-scale reactions and tests, e.g. qualitative analysis (p. 135) or solvent
selection for recrystallization (p. 93).

Beakers
Beakers are used for general purposes, e.g. heating a non-volatile solvent
while the solute dissolves, 'working up' a reaction where liquid/solid
products need to be accessible for manipulation (stirring with a glass rod),
or titrations using electrodes where a wide opening is essential (see p. 229).
The volume graduations on the side are inaccurate and should only be used
where approximations will suffice. The lip on the beaker is specifically
designed to aid quantitative transfer of solutions (see p. 18).
Conical (Erlenmeyer) flasks

These can be used for general purposes, but they have more specialist
applications. The narrow mouth and sloping shoulders reduce losses on
stirring/swirling and evaporation and make them easier to seal. The absence
of a lip does not favour quantitative transfer: useful in manual titrations
(p. 145) and recrystallizations (p. 96). Volume markings are approximate.
Bottles and vials

These can be used when the solution needs to be sealed for safety, storage
or to prevent evaporation or oxidation. They usually have a screwtop,
plastic cap or stopper or a ground-glass stopper and come in various styles
and sizes: from 2.5 L glass bottles used for storing large volumes of
solutions to small plastic-capped vials (5mL) for saving small amounts of
reaction products.
Seal the vessels in an appropriate manner, using a stopper, cap or sealing
film such as Parafilm'f or Nescofilm'P, bearing in mind the nature of the
contents - sealing film should only be used for water solutions since it
dissolves in some organic solvents and plasticizers may be extracted. Do not
use corks, they are not air-tight. Do not use rubber bungs to seal containers
containing organic solvents, they can swell even over a short period of time
making removal very difficult.
You should clearly label all stored solutions (see p. 21) including all
relevant hazard information.
Creating specialized apparatus

Glassware systems incorporating ground-glass connections, such as
Quickfit'P, are essential for setting up combinations of standard glass
components for reactions, distillation, etc. In project work you may need to
adapt standard forms of glassware for a special need. It may be necessary
to contact a glass blowing service to make special items to order.
Table 3.2 Spectral cutoff values for glass and

plastics (A50 = wavelength at which
transmission of electromagnetic radiation is Choosing between glass and plastic containers
reduced to 50%)
Bear in mind the following points:
• Reactivity. Plastic vessels often distort at relatively low temperature, may
Routine glassware 340 be inflammable, may dissolve in certain organic solvents and may be
Pyrex® glassware 292 affected by prolonged exposure to ultraviolet (UV) light.
Polycarbonate 396 • Opacity. Both glass and plastic absorb light in the UV range of the
Acrylic 342
Polyester 318
electromagnetic spectrum (Table 3.2). Quartz should be used where this is
Quartz 220 important, e.g. in cells for UV spectrophotometry (see p. 165) or
photochemistry.

• Contamination. Some plasticizers may leach from vessels, especially with

some organic solvents, such as DCM. Glass may adsorb ions and other
molecules and then leach them into solutions, especially under acidic or
alkaline conditions. Pyrex@ glass is stronger than ordinary soda glass
(rarely found except in specific items such as Pasteur pipettes and melting
point tubes, but check if you are not sure) and can withstand
temperatures up to 500°C.
• Rigidity and resilience. Plastic vessels are not recommended where
volume is critical as they may distort through time. Glass vessels are more
easily broken than plastic.
• Disposability. Plastic items may be cheap enough to make them
disposable, an advantage where there is a risk of chemical contamination.
Cleaning glass and plastic

In quantitative analytical work, beware of contamination arising from prior
use of chemicals or inadequate rinsing following washing. A thorough rinse
with distilled or deionized water immediately before use will remove dust
and other soluble deposits, but ensure that the rinsing solution is not left in
the vessel. For analyses on the 'f1.g scale' and below, you should clean glass
and plastic containers by immersion in nitric acid (10% v/v) overnight and
then rinsing with a large volume of distilled or deionized water. The clean
vessels should be stored either upside down or covered with Clingfilm'P, to
prevent dust contamination.
For general work, 'strong' basic detergents (e.g. Decon® or Pyroneg'P)
are good for solubilizing acidic deposits and an acid wash will remove
remaining basic residues. A rinse with ethanol or propanone (acetone) will
remove many organic deposits.
Safety with glass

Many minor accidents in the laboratory are due to lack of care with
glassware. You should follow these general precautions:
• Wear eye protection at all times.

• Don't use chipped or cracked glassware and examine the equipment for
'star' cracks - it may break under very slight strains and should be disposed
of in the broken glassware bin. All laboratories will have a waste bin
dedicated to broken glass. Never put broken glass into other bins.
• If heating glassware, use a 'soft' Bunsen flame (half-open air vent) or
'wave' the flame around the heating point - this avoids creating a hot
spot where cracks may start. Always use special heat-resistant gloves or
rubber 'fingers' (see p. 36) when handling hot glassware.
• When clamping glassware (see p. 26) ensure that the clamp has a cork,
rubber or plastic 'cushion' in the jaws to prevent breakages. There must
be no metal-glass contact and you must not over-tighten the clamp.
• Take care when attaching rubber or plastic tubing to glass tubes,
condensers, etc., and inserting thermometers and glass tubes into
screwcap adapters (see p. 43). Always hold the tube and the 'hole' close
together (Fig. 3.5) and wear thick gloves where appropriate.
• Don't force bungs too firmly into bottles (see p. 12) they can be difficult
to remove. If you need a tight seal, use a screwtop bottle, with a rubber
Fig. 3.5 Handling glass tubing. or plastic seal, Parafilm'P or ground-glass jointware, such as Quickfit'P.

• Never carry large bottles (> 1L) by their necks - carry them in a bottle
basket.
• Dispose of broken glass thoroughly and with great care - use disposable
paper towels, tongs or dust-pan and brush and thick gloves. Always put
pieces of broken glass in the correct bin.

------~8 Basic laboratory procedures I
Using chemicals
Safety aspects
In practical classes, the person in charge has the responsibility to inform you
of any hazards associated with the use of chemicals. In project work, your
first duty when using an unfamiliar chemical is to find out about its pro-
perties, especially those relating to safety. For routine practical procedures, a
risk assessment may have been carried out by a member of staff and the
relevant safety information may be included in the practical schedule: an
example is shown in Table 4.1. If not, you will have to carry out a risk
assessment before you begin. Before you use any of the chemicals, you must
find out whether safety precautions need to be taken and complete the
appropriate forms confirming that you appreciate the risks involved. Your
Table 4. 1 Representative risk assessment
information for a practical exercise in organic
department must provide the relevant information to allow you to do this. If
chemistry: the synthesis of N-phenylethanamide your supervisor has filled out the form, read it carefully before signing.
(acetanilide) Key safety points when handling chemicals are:
• Treat all chemicals as potentially dangerous.

• Wear a lab coat, with the buttons fastened, at all times.
Aminobenzene Toxic, harmful by skin • Wear eye protection at all times.
absorption. Wear gloves,
dispense in fume cupboard • Make sure you know where safety devices such as eye bath, fire
extinguisher, first aid kit are kept before you start work in the lab.
Ethanoic Corrosive, flammable, toxic,
anhydride reacts with water. • Wear gloves for toxic, irritant or corrosive chemicals and carry out
Wear gloves, dispense in procedures with them in the fume cupboard.
fume cupboard • Use appropriate aids such as spatulae, pipette fillers, Pasteur pipettes etc.,
to minimize risk of contact.
• Label all solutions, samples, etc., appropriately (see p. 22).
• Extinguish all naked flames when working with flammable substances.
• Never drink, eat or smoke where chemicals are being handled.
• Report all spillages and clean them up appropriately.
• Dispose of chemicals in the correct manner.
Selection
Chemicals are supplied in varying degrees of purity and this is always stated
on the manufacturer's containers. Suppliers differ in the names given to the
grades and there is no conformity in purity standards. Very pure chemicals
cost more, often very much more, and should only be used when the
situation demands. If you need to order a chemical, your department will
have a defined procedure for doing this.
Preparing solutions
Solutions are usually prepared with respect to their molar concentrations (e.g.
mot l.:', mol dm ", or mol m r or mass concentrations (e.g. gL-I, or
')
kg rn"): both can be regarded as an amount (usually mass) per unit volume,
in accordance with the relationship:
. amount
concentration = I [4.1]
vo ume

Basic laboratory procedures I
The most important aspect of eqn [4.1] is to recognize clearly the units
involved, and to prepare the solution accordingly: for molar concentrations
you will need the relative molecular mass of the chemical, so that you can
calculate the mass of substance required. Further advice on concentrations
and interconversion of units is given on p. 45.
In general there are two levels of accuracy required for the preparation of
solutions:
1. General-purpose solutions - solutions of chemicals used in qualitative
and preparative procedures (p. 17) when the concentration of the
chemical need not be known to more than one or two decimal places.
For example:
(a) solutions used in extraction and washing processes, e.g. hydrochloric
acid (0.lmoIL-1), sodium hydroxide (2M), sodium carbonate (5%
w/v);
(b) solutions of chemicals used in preparative experiments where the
techniques of purification - distillation, recrystallization, filtration,
etc. - introduce intrinsic losses of substances that make accuracy to
any greater level meaningless.
2. Analytical solutions - solutions used in quantitative analytical proce-
dures when the concentration needs to be known to an accuracy of four
decimal places (e.g. 0.0001 mol L -1). For example in:
(a) volumetric procedures (titrations) and gravimetric analysis, where
the concentrations of standard solutions of reagents and compounds
to be analysed need to be accurately known;
(b) spectroscopy, e.g. quantitative UV and visible spectroscopy, atomic
absorption spectroscopy and flame photometry;
(c) electrochemical measurements: pH titrations, conductance measure-
ments and polarography;
(d) chromatographic methods.
The procedures for weighing and the glassware used in the preparation of
solutions differ according to the level of accuracy required.
Preparation of general-purpose solutions

Box 4.1 shows the steps involved in making up general-purpose aqueous
solutions.
The concentration you require is likely to be defined by the protocol you are
following and the grade of chemical and supplier may also be specified. To
avoid waste, think carefully about the volume of solution you require, though it
is always advisable to err on the high side because you may spill some, make a
mistake when dispensing or need to repeat part of the experiment. Choose one
of the standard volumes for vessels, as this will make measuring out easier.
Use distilled or deionized water to make up aqueous solutions and stir
with a clean Pyrex® glass rod or magnetic stirrer bar ('flea') until all the
chemical is dissolved. Magnetic stirrers are a convenient means of stirring
solutions but precautions should be taken to prevent losses by splashing. Add
the flea to the empty beaker or conical flask, add the chemical and then some
water. Place the vessel centrally on the stirrer plate, switch on the stirrer and
gradually increase the speed of stirring. When the crystals or powder have
dissolved, switch off the stirrer and remove the flea with a magnet. Take care
not to contaminate your solution when you do this and rinse the flea into the
solution with distilled water. In general it is convenient to use glass rods with

beakers - ease of access for stirring - and magnetic fleas with conical flasks -
lower losses through splashing - but often it is a matter of your preference
and laboratory skills.
'Obstinate' solutions may require heating, but only do this if you know
that the chemical will not be damaged at the temperature used. Use a stirrer-

heater to keep the solution mixed as you heat it. Allow the solution to cool
to room temperature before you finalize its volume.
Preparation of analytical solutions

The key features in the preparation of solutions for analytical purposes are:
• Make sure that you have the most accurate available knowledge of
masses of the chemicals used.
• Ensure that you have the most accurate available knowledge of the
volumes of solutions used.
To achieve these features exact techniques of weighing and solution transfer
must be used and the procedure is illustrated in the following example.
'Prepare a standard solution (250.00 mL) of ammonium ferrous
sulphate (approximately 0.1 M), which is to be used to determine the
concentration of a solution of potassium permanganate by titration'.
You must be aware of the following embedded information:

• This is a quantitative experiment, therefore requiring an analytical solution
to be prepared.
• You must use a 250.00 mL volumetric flask, which you should note is
calibrated at 20°C.
• You must weigh accurately, to four decimal places, the mass of the
chemical.
• It is almost impossible to weigh the exact mass of chemical for a specific
concentration. For example, the mass of (NH4hFeS04.6H20 required to
prepare 250.00mL of 0.1 M solution is 9.8035g and you cannot weigh
out this exact mass. However, you can weigh out a known mass to four
decimal places accurately. From this you can then calculate the exact
concentration of the chemical in solution, since you will know both mass
and volume to a high degree of accuracy.
Box 4.2 shows the method for the preparation of the standard solution.
The main practical point is that you must not lose, by splashing or failure to
transfer by inadequate rinsing, any of the solution being prepared in the
beaker and you must transfer all of the solution, by repeated rinsing, into the
volumetric flask. Therefore it is good practice to use only a glass rod to stir
the solution gently to dissolve the solid and to use the glass rod, as shown in
Fig. 4.1, to pour the solution into the filter funnel. This technique, with
practice, prevents losses of solution down the side of the beaker via the
spout; rinsing with water can be achieved by use of a wash bottle squirted
directly into the beaker.
You should not use a flea to stir a solution in the preparation of a
standard solution, since this introduces more washing steps - washing the flea
and the 'flea extractor' - and you still need to use the glass rod for
Fig. 4.1 Pouring a solution using a glass rod. quantitative transfer.
Procedure required with analytical solutions prepared from liquids

Many experiments in analytical chemistry, such as chromatography and
spectroscopy, require the preparation of a standard solution of a liquid
organic compound. Therefore you must know accurately the mass of the
liquid. The compound can be dispensed by the methods described in Chapter
3, provided that the pipette, syringe, etc., is accurate, and thus the mass =
volume x density, bearing in mind the temperature factor.

Basic' laboratory procedures I
Alternatively you can use a weighing bottle as shown in Fig. 4.2. The
liquid is placed in the bottle, weighed accurately and then the approximate
amount required is added to the volumetric flask containing some solvent.
The volumetric flask is stoppered immediately, the weighing bottle reweighed
and the weight of liquid dispensed is calculated. The volumetric flask is then

made up to the mark and stoppered. You now know the concentration of the
teat
standard solution to four decimal places.
Pasteur pipette Preparing dilutions

Making a single dilution
In analytical work, you may need to dilute a standard solution to give a
screwcap adapter particular mass concentration or molar concentration. Use the following
procedure.
1. Transfer an appropriate volume of standard solution to a volumetric
flask, using appropriate equipment (Table 3.1).
2. Make up to the calibration mark with solvent - add the last few drops
5mL Quickfit® conical flask from a Pasteur pipette until the bottom of the meniscus is level with the
calibration mark.
3. Mix thoroughly, either by repeated inversion (holding the stopper firmly)
Fig. 4.2 A weighing bottle. or by prolonged stirring, using a magnetic stirrer. Make sure that you
add the magnetic flea after the volume adjustment step.
For general-purpose work using dilute aqueous solutions where the higher
degree of accuracy is not required, it may be acceptable to substitute conical
flasks, beakers or test tubes for volumetric flasks and use measuring cylinders
for volume measurements. In such cases you would calculate the volumes of
'stock' solutions (usually 'bench' reagents) and diluent required, with the
assumption that the final volume is determined by the individual volumes of
stock solution and diluent used. Thus a two-fold dilution would be prepared
by using one volume of stock solution and one volume of diluent. The dilution
factor is obtained from the initial concentration of the stock solution and the
fmal concentration of the diluted solution. The dilution factor can be used to
calculate the volumes and stock and diluent required in a particular instance.
For example, suppose you wanted to prepare 100mL of a solution of NaOH
at 0.lmolL-1. Using the bench reagent, commonly containing 2.0molL-]
(2.0 M), the dilution factor is 0.1 -i- 2.0 = 0.05 = 1/20 (a twenty-fold dilution).
Therefore the amount of stock solution required is 1/20th of 100mL = 5mL
and the amount of diluent needed is 19/20th of 100mL = 95 mL.
Preparing a dilution series

Dilution series are used in a wide range of procedures including the
preparation of standard curves for the calibration of analytical instruments
(p. 171). A variety of different approaches can be used but the most common
is a linear dilution series.
In a linear dilution series the concentrations are separated by an equal
amount, e.g. a series containing cadmium at 0, 0.2, 0.4, 0.6, 0.8, 1.0mmol L -\
might be used to prepare a calibration curve for atomic absorption spectro-
scopy (p. 170) when assaying polluted soil samples. Use [Cd VI = [C2]V2 to
calculate the volume of standard solution for each member of the series and
pipette or syringe the calculated volume into an appropriately sized volumetric
flask as described above. Remember to label clearly each diluted solution as you
prepare it, since it is easy to get confused. The process is outlined in Box 4.3.
~ Make all the dilutions from the working stock solu-

tion to the required solution. Do not make a solution of lower dilution
from one already prepared: if you have made an error in the first dilution,
it will be repeated for the second dilution.

Storing chemicals and solutions

Chemicals which decompose easily (labile chemicals) may be stored in a
fridge or freezer. Take special care when using chemicals which have been
stored at low temperature: the container and its contents must be allowed to
warm up to room temperature before use, otherwise water will condense onto
the chemical. This may render accurate weighing impossible and you may
ruin the chemical.
Chemicals and solutions to be stored at low temperatures must be in
stoppered or sealed vessels. Do not store aqueous solutions below 0 QC since
freezing can occur and, with the resulting expansion of the volume, the vessel
may crack. Solutions containing flammable solvents should only be stored in
specialized 'spark-proof fridges: consult your laboratory instructor.
You must be aware of the particular problems of storing solutions in flasks
with ground-glass joints. If you are using aqueous solutions you should

lightly grease the joint and stopper with petroleum jelly, since the water will
not dissolve the grease as it is poured from the flask. The stopper can be
removed easily and the solution will be uncontaminated.
Conversely, if you are using solutions made up from organic solvents, you
should not grease the joints since the organic solvent will dissolve the grease
as you pour it from the flask and contaminate the solution. Moreover, you
should not allow the solution to come in contact with the un greased joints,
since the solvent will evaporate and leave the solute to 'weld' the stopper to
the flask. Fill the flask with solution, using a filter funnel with the stem of the
funnel positioned well below the joint. See p. 43 for use of ground-glass
jointware.
~ Always label all stored chemicals clearly with the fol-

lowing information: the name (if a solution, state solute(s) and concen-
tratlonlsl], plus any relevant hazard warning information, the date made
up, and your name.
Balances and weighing

Electronic single-pan balances with digital readouts are now favoured over
mechanical types and are common in most laboratories. There are essentially
two types of balance:
1. General purpose balances which weigh to the nearest 0.01 g with a
capacity of about 300 g. Chemicals may be dispensed for weighing, into a
suitable weighing container, directly onto these balances.
2. Analytical four-figure balances for quantitative work, which weigh to
the nearest 0.0001 g (0.1 mg) and have a maximum capacity of about
100 g. Chemicals must not be transferred onto the balance at any
time and analytical balances must only be used for weighing by
difference.
Both types are illustrated in Fig. 4.3 and you should familiarize yourself with
their operation before use.
General-purpose balances
The most useful feature of this type of balance is the electronic zero facility
(self-taring), which means the mass of the weighing container can be
subtracted automatically before weighing chemicals.
To operate a standard self-taring balance:
1. Check that it is level, using the adjustable feet to centre the bubble in the
spirit level (usually at the back of the machine). For relatively accurate
work or when using in a fume cupboard, make sure that the draught
shield is in place.
2. Ensure that the balance is switched on: the display should be lit.
3. Place an empty weighing container (see p. 24) centrally on the balance
pan and allow the reading to stabilize. If the object is larger than the pan,
take care that no part rests on the body of the balance or the draught
shield as this will invalidate the reading. Press the tare bar to bring the
reading to zero.
4. Place the chemical or object carefully in the weighing vessel:
(a) Solid chemicals should be dispensed with a suitably sized clean
spatula.

(b) Non-volatile liquids should be dispensed using a Pasteur pipette but

take the weighing container off the balance pan before dispensing;
then reweigh the liquid plus container. Repeat until the desired
weight is obtained.
5. Allow the reading to stabilize and make a note of the reading.
6. If you have added excess chemical, take great care when removing it.
Remove the container from the balance, remove the solid (with a
spatula) or liquid (with a Pasteur pipette) and reweigh.
7. If you need to clean any deposit accidentally left on or around the
balance, switch off the balance.
Take care not to exceed the limits for the balance: while most have devices to
protect against overloading, you may damage the mechanism.
Analytical balances
These are delicate precision instruments and as such are likely to be found
away from the open laboratory in draught-free conditions on a vibration-
dampened surface. Analytical balances are maintained to the highest
specifications and should need no adjustments on your part, such as levelling
and zero adjustment. The key points for using an analytical balance are
summarized below:
• No chemicals must be transferred within the weighing compartment of

the balance.
• If it has a 'locking' function, the balance pan must always be 'locked'
when placing and removing objects onto and from the balance pan.
• The doors of the balance must always be closed when taking measure-
ments.
The procedure for weighing a solid chemical for the preparation of an

analytical standard solution is shown in Box 4.4.
(a)
Weighing containers
These come in various materials, shapes and sizes: from glass weighing boats
to beakers and even special glazed paper. The weighing container to be used
depends on several factors:
• The amount of chemical to be weighed.

• The properties of the chemical to be weighed: is it solid, liquid, volatile,
corrosive, deliquescent, hygroscopic?
• How and into what type of vessel it is to be transferred.
• The accuracy to which it is to be weighed.
Some of the common types of weighing container are shown in Fig. 4.4.
For analytical procedures, only weighing boats, weighing bottles or glass
or plastic sample tubes should be used. Weighing boats are used to transfer a
solid directly into a volumetric flask via the neck of the weighing boat: this
procedure is recommended when the chemical is known to be totally soluble
in the solvent and allows you to omit the solution preparation stage in a
beaker or conical flask (see Box 4.2). You must ensure that the neck of the
(b) weighing boat will fit well inside the ground-glass joint of the neck of the
Fig. 4.3 Single-pan balance: (a) general
volumetric flask so that all the chemical can be washed down the sides of the
purpose, two decimal places; (b) analytical, four volumetric flask and does not stick to the ground-glass joint during the
decimal places. quantitative transfer.

(a)
(b)
(c)
Fig. 4.4 Weighing containers: (a) plastic

weighing dishes; (b) weighing bottles; (c)
weighing boat.
For general-purpose work, weighings can be made directly into pre-

weighed or 'tared' conical flasks or beakers, again to avoid a transfer stage.
Much more common is the use of disposable plastic weighing dishes of the
appropriate size. The edges of these dishes can be squeezed together to form
a 'funnel' to prevent losses when transferring the solid. Remember that
plastic disposable dishes may dissolve in organic solvents such as prop an one
(acetone), toluene, etc., and should not be used for low-melting organic solids
or liquids. Watch- and clock-glasses should be avoided if you wish to transfer
the solid into narrow-necked vessels such as conical flasks or sample tubes
since it very difficult to direct the solid into the narrow opening of the vessel
from the large 'flat' surface of the watch- or clock-glass. In such cases, or
Fig. 4.5 Transferring a solid using glazed when large amounts of solid are to be transferred, it is advisable to use a
paper. wide-necked filter funnel called a 'powder funnel'.

In many preparative experiments, which are carried out on a small scale

(involving I g to 10g of solids), the most useful weighing container is special
glazed paper, provided that the chemicals do not react with the paper. A
folded glazed paper creased square of glazed paper is 'tared' on the balance pan and the solid
weighed out directly onto it. The chemical can then be allowed to flow down
solid
the crease into the vessel (Fig. 4.5). Furthermore, when attempting to transfer
rolled glazed paper small amounts of solid in vessels with narrow-bore ground-glass joints (see
p. 43) it is important not to allow the solid to contact the joint, because the
joint will not seal correctly. Use a filter funnel or roll a piece of glazed paper
into a funnel, insert the stem of the paper funnel to below the joint and then
run in the solid from the creased weighing paper (Fig. 4.6). Paper used in this
manner is much cheaper than proprietary weighing dishes and is a useful
flask
method of recycling out-of-date manufacturers' catalogues!
Fig. 4.6 Transferring a solid to a narrow-

necked flask.

-------8 Basic laboratory procedures 11
Clamps and support stands

When carrying out experiments it is vital that all apparatus is held in place
securely during the procedure. It is essential that you know how to assemble
supporting and securing equipment to the highest possible standards of
safety. The most common types of supporting and securing equipment are
shown in Fig. 5.1.
Support stands
These are also known as 'retort stands' and comprise an aluminium or steel
rod screwed into a heavy metal base. Always check that the base sits level
and that the rod is tightened fully in place.
Clamp holders
support stand
These are also described as 'clamp bosses' or 'bosses'. Make sure that the
locking screws move freely and are not distorted. When you attach the clamp
holder to the support stand, tighten the screw firmly and ensure that the open
'slot' to be used for the clamp is pointing upwards (Fig. 5.2).
Clamps
General-purpose clamps are used for securing glassware - therefore make
three-finger clamp clamp holder sure that the inner surfaces of the clamp 'jaws' and the 'fingers' are covered
with cork or rubber to provide a cushion for the glass: there must be no
metal to glass contact in case you overtighten the clamp and crush the glass.
Tighten the clamp firmly and ensure that the clamped glassware does not
- - .0/.·,',"','.'.·,' . move.
~._o O.0
. ~
..
--"-,-=
....." ....-..
Conical flasks should be clamped at the neck and ground-glass jointware
~ ~
should be clamped at the joint - this usually has the greatest thickness of
clamp metal ring
glass.
~ Take particular care when using parallel-sided separa-

tory funnels and chromatography columns (Fig. 5.31, where clamping in
the middle of the funnel can be the same as squeezing the middle of a
large-diameter glass tube. Clamp at the ground-glass joint using a 'well-
cushioned' clamp.
In most clamps, only one of the jaws moves when turning the screw. When
burette clamp
you use the clamp in a horizontal position, make sure that the movable jaw is
Fig. 5.1 Clamps and supporting equipment. at the top (see p. 109).
Burette clamps are specially designed to hold the burette vertically. Springs
hold the burette at two points about 5 cm apart - again check for the
presence of a rubber or plastic 'cushion' at the points of contact - to prevent
slipping and lateral movement. Since the burette clamp slides down the rod
of the support stand, then provided the support stand is vertical, the burette
will be vertical.
Support rings
These metal rings come in various diameters to support filter funnels and
separatory funnels. Often these support rings are coated in plastic to provide
the cushion between metal and glass.

Basic laboratory procedures 11
If your support ring is metal, you can make a 'cushion' by finding a piece
of thin-walled rubber tubing of the same bore as the metal of the ring,
cutting it to a length equivalent to the circumference of the ring and then
cutting down the length of the rubber tubing. You can then slide the tubing
around the ring to provide the 'cushion'.
RIGHT WRONG
~ When using clamps, support rings and support stands

movable jaw on TOP make sure that the clamped/supported apparatus is always in position
above the base of the support stand (Fig. 5.4) to prevent the stand top-
pling over.
~
t
fixed jaw underneath to
Cork rings
WRONG
support, e.g. a condenser These are used to hold round-bottom flasks on flat surfaces while
manipulations are being carried out. Since they are light in weight they can
Fig. 5.2 Right and wrong ways of using
be used to hold round-bottom flasks on a general-purpose balance.
support stands, clamp holders and clamps.
Filtration
Filtration is the physical separation of a solid from a liquid and is a process
encountered in experimental procedures such as gravimetric analysis (p. 139),
recrystallization (p. 92), and solvent drying (p. 41). In principle, the mixture
of the solid and liquid is passed through a porous material, filter paper or
sintered glass, and the solid is trapped on the porous material while the liquid
passes through.
The type of filtration equipment you select for use depends upon which of
the two components, the solid or the liquid, you are trying to isolate. In
general:
• If you wish to isolate the liquid - use gravity filtration.
• If you wish to isolate the solid - use suction (vacuum) filtration
Gravity filtration
In gravity filtration you need to pass the liquid through the porous material
and retain all the unwanted solid in the filter. In general, the best material to
use is a filter paper of the appropriate porosity to trap all the solid particles
and with the greatest surface area to allow the liquid to pass through quickly.
The apparatus required for gravity filtration is shown in Fig. 5.5. The filter
funnels are usually made of glass, but if organic solvents are not involved in
the filtration, plastic funnels can be used. Glass filter funnels with the pipe
cut off are known as 'stemless' filter funnels and have a specific use in hot
filtration (p. 98).
The key to successful gravity filtration is the fluted filter paper. A fluted
filter paper decreases the area of contact between the filter paper and the
funnel, thus allowing rapid filtration. If you use 'traditional' cone-folded filter
paper, note that all sides of the paper are touching the sides of the funnel and
on half the filter paper the liquid has to pass through three thicknesses of
paper, all of which slow the rate of filtration. Slow filtration can lead to
disaster in hot filtration during recrystallization (p. lOO).
Since filter funnels and filter papers come in different sizes, choose a filter
paper of diameter just less than twice the diameter of the funnel. When
fluted, the filter paper will be just below the rim of the funnel. There are
RIGHT
many ways to fold (flute) a filter paper, but one of the simplest is shown in
Fig. 5.3 Clamping a parallel-sided funnel. Box 5.1.

RIGHT WRONG
Fig. 5.4 Correct use of support stands.
Fig. 5.5 Gravity filtration.
To filter the mixture, swirl the suspension of the solid in the liquid so that
there is a fairly even distribution of solid in the liquid, and then pour the
mixture into the filter cone, making sure that you do not pour any of the
mixture outside the filter paper otherwise you will need to repeat the
filtration, and do not overfill the filter cone. Transfer all the mixture in this
way and finally wash the last bit of solid and liquid into the filter cone with a
small amount of filtered solution and then a little pure solvent.
Suction filtration
This technique is used for the isolation of a solid from a suspension of a solid
in a liquid and relies on producing a partial vacuum in the receiving flask.
The essential components of a suction filtration system are:
• A ceramic funnel containing a flat perforated plate: there are two types
based on size and shape called Buchner funnels or Hirsch funnels. When
you are filtering, the perforated plate is covered by a filter paper.

• A receiver flask with a side arm for attachment of the vacuum source.
Buchner flasks are conical flasks made from thickened glass, and Hirsch
tubes (also known as side-arm boiling tubes or test tubes depending upon
size) are capable of withstanding weak vacuum, e.g. a water pump.
• A flexible seal between the ceramic funnel and the receiving vessel: a
Buchner collar or filter seal.
• A source of vacuum, usually a water pump (water aspirator) which is
connected to the receiving flask by thick walled rubber tubing (pressure
tubing). Sometimes there will be a trap between the water pump and the
recei ving flask.
The various types of these components are shown in Fig. 5.6: typical
apparatus is shown in Fig. 5.7 and the general procedure for suction
filtration is described in Box 5.2.
Buchner funnel Hirsch funnel
side-arm
Buchner flask
boiling tube
or test tube
Fig. 5.6 Equipment for suction filtration.
thick-walled tubing
flat filter paper vacuum release tap
Neaprene®
adapter
clamp
thick-walled filter flask

(Buchner flask)
Fig.5.7 Suction filtration using a Buchner funnel.


Instead of a filter paper on a porous plate, sintered-glass can be used as the

porous material for filtration. Sintered-glass funnels come in various types
and sizes (Fig. 5.8) with different porosity (size of holes) of the sintered glass.
Sintered glass is also used in crucibles (Fig. 5.8) in gravimetric analysis
(p. 139).
sintered-glass vacuum take-off
filter disk side-arm
.'i""'OO'"'~!Pj
filter ~~:ss ~
standard
sinter for
sinters for taper joint
gravimetric
filtration
analysis
Fig. 5.8 Typical sintered-glass funnels.
When sintered-glass funnels are used instead of Buchner or Hirsch funnels

the solid is collected directly on the sintered glass: no filter paper is used. As
a result cleaning sintered-glass funnels is a major problem if the solid has
been drawn into the pores of the sintered-glass. Therefore if you are to use a
sintered glass funnel, check with your instructor on the appropriate method
for cleaning the funnel, before and after use, so that your product will not be
contaminated. On the other hand, if particle sizes are large enough to prevent
this problem, sintered-glass funnels are very effective in suction filtration.
Heating
In the laboratory you will be required to heat chemicals in dissolution of a
solid, promotion of reaction (reflux), distillation of pure compounds and
mixtures, extraction, coagulation of precipitates, drying solid compounds, etc.
Your choice of heat source depends upon several factors:
• First and foremost, the flammability and volatility of the chemical and
solvent.
• The operation to be carried out, e.g. simple preparation of a solution,
reflux or distillation.
• The temperature required for the process.
• The amount of chemical or solvent to be heated.
Bumping
Before you attempt to heat any liquid or solution you must take precautions
to prevent 'bumping'. This is when the liquid suddenly boils without any
warning and results in hot liquid and vapour shooting uncontrollably out of
the container. 'Bumping' can occur during simple heating in a test tube,
conical flask or beaker or in more complex situations such as reflux and
distillation. It is necessary to provide a point in the liquid or solution where
vaporization of the liquid can occur in a controlled manner.
Before you start heating a liquid or solution you must always take one of
the following 'anti-bumping' precautions:
• Add one or two 'boiling stones' or 'anti-bumping granules'; these can be

filtered off later in the process.

hot blue flame

when air inlet open
luminous wavy flame

when air inlet closed
(a) (b)
Fig. 5.9 (a) Bunsen burner; (b) microburner.
• Add a pyrex® glass rod to the beaker or conical flask. The rod must be
longer than the container so that it can be removed and rinsed before the
solution is used further.
• Add a 'boiling stick': these are thin pieces of wood sold as 'wooden
applicators', but you must be sure that nothing will be extracted from the
wood into your solution.
• Stir the liquid or solution with a magnetic 'flea' (see p. 35) which should
be removed before further processing of the chemicals.
• Stir the liquid or solution with a mechanical stirrer (see p. 118).
• Use an air or inert gas (nitrogen) capillary bleed during vacuum distillation
(see p. 110).
~ Make sure that you add the anti-bumping device

before you start heating since uncontrolled boiling may occur immedi-
ately. You must allow the liquid to cool back to room temperature and
then add the anti-bumping device.
Burners
Gas burners come in two common forms: large burners called Bunsen
burners and small burners known as microburners (Fig. 5.9). Bunsen burners
are commonly used for heating aqueous solutions in flat-bottomed pyrex®
vessels supported on a tripod and wire gauze (Fig. 5.10) but for many other
heating applications Bunsen burners do not provide adequate control.
Microburners may be used for direct heating of round-bottom or pear-
shaped glassware in small-scale operations where good control of the heating
rate is required, such as distillation or determination of melting point (see
p. 87). When using a microburner for heating make sure that you do not
create a 'hot spot', which may result in uneven heating of the liquid and
'bumping', by 'waving' the flame around the flask starting just below the
level of the liquid and working down to the bottom of the flask and back
again.
Microburners are also useful for heating boiling tubes and test tubes,
sealing the ends of melting point tubes (see p. 88), making micropipettes for
chromatography (see p. 217) and bending the ends of Pasteur pipettes for
special purposes (see p. 112).
Fig. 5.10 Heating an aqueous solution using a To use a burner, first make sure that the gas piping, is attached
Bunsen burner. securely to both the burner and the gas tap, is of the correct type of

rubber tubing, and is not damaged. Close the movable collar and light the
gas with a gas igniter: do not use a splint or a piece of burning paper in
case you cause a fire in the waste bin when disposing of the paper. Open
the collar to produce a hot blue flame and adjust the gas flow to give the
required size of flame. If your work is interrupted, close the collar a little
to produce a luminous flame. Finally, when the operations are complete,
turn off the gas and do not pick up the burner by the barrel, or put it
into a cupboard, until it has cooled.
The advantages of the burner are that the heat source can be removed
instantly from the apparatus and that the flame is visible: in contrast you
often cannot distinguish between a cold and hot metal surface. The
disadvantages are those of an open flame.
Steam baths and water baths

These are heat sources, which have a maximum temperature of 100°C; they
are safe to use even with most flammable chemicals and solvents and they
differ only in the way the steam is produced.
Water baths are the more common of the two, and the steam is
generated by heating water with an electric element - just like a kettle. The
element may have a thermostatic control, which can control the
temperature of the water to some extent. Most water baths have a constant
level device on the side of the bath, which supplies water to the bath to a
fixed level above the heating element and prevents the water bath from
boiling dry. Water baths are 'single hole' or 'multiple hole' types, and the
holes are covered by concentric metal or plastic rings, which allows you to
vary the size of the hole.
When you are to use a water bath, make sure that water is flowing into
and out of the bath via the constant level device - check that water is flowing
from the pipe into the drain. Turn on the controller to the level you require -
the power can be turned down once the bath is boiling.
A steam bath looks like the 'single hole' water bath except that there is no
heating element and no constant level device. Steam baths require piped
steam as their source of heat usually provided by a steam line, which is a
permanent supply in the laboratory.
Beakers and conical flasks sit firmly on the rings, while round-bottom and
pear-shaped flasks should have about half the surface of the flask immersed
in the steam (Fig. 5.11).
The main advantage of a water bath is the minimal risk of fire, while the
major disadvantages are the maximum temperature available (~100°C)
and the special precautions needed if anhydrous reaction conditions are
required: remember that steam will condense down the inside of reflux
condensers.
Hot plates and stirrer hot plates

These consist of a flat metal or ceramic plate, which is heated electrically, and
varies in size for use by an individual or by several people at the same time.
The small versions normally have a built-in magnetic stirrer, which can be
used to stir the liquid with a magnetic 'flea'.
Hot plates of both types should only be used for heating flat-bottom
vessels such as beakers or conical flasks and only when the liquid being
heated is non-flammable. The vapour of a flammable liquid may 'run' down
the outside of the vessel and ignite on the hot metal of the heating surface.
Since the contact area between the heating surface and a round-bottom flask

water out
./
water in
water out
~
~
-
water in water out
~
water in
(a) (h)
Fig. 5.11 (a) Heating a conical flask on a water bath; (b) heating a reflux set-up on
a water bath.
is very small, your attempts to heat these flasks effectively will require
excessive temperatures of the heating surface, which increases the dangers due
to lack of control of the heating process. Therefore round-bottom flasks
should only be heated using an oil bath (below) or a mantle (p. 35).
The flat exposed heating plate is extremely dangerous when hot: always
check that the plate is cool by passing your hand over the plate without
touching it or by placing a drop of water on the plate. If you have to pick up
a hot plate, hold it by the sides and do not touch the plate; it may burn.
Typical uses of hot plates are illustrated in Fig. 5.12a.
Oil baths
These are mostly used to heat round-bottom flasks at temperatures above
100°C. The oil bath, containing the heating fluid, is usually a non-ferrous
metal or pyrex® dish and heated on a stirrer hot plate, and the temperature
of the bath is measured with a thermometer. The oil bath should never be
more than half-full, to allow a margin of safety for thermal expansion of the
oil, and stirred with a magnetic 'flea' to ensure even heating. The equipment
used in a typical oil bath is shown in Fig. 5.12b.
The nature of the oil used in the bath depends upon the temperature range
required and a selection of liquids is shown in Table 5.l.
You are most likely to encounter paraffin oil baths during your laboratory
work and the following safety points should be considered:
• Paraffin oil discolours rapidly with prolonged heating. If the oil is dark,
replace it with fresh oil. Dispose of the old oil in an appropriate manner
(check with laboratory staff).
• Check that there is no water in the bottom of the oil bath: look for a
separate layer or globules of liquid in the bottom of the bath. If you heat

Table 5.1 Oil bath liquids

clamp
Paraffin oil 0-200 Flammable, cheap, produces acrid

(mineral oil) smoke above 220°C
Ethylene glycol 0-150 Flammable, cheap, low flash point
Polyethylene glycol 400 0-250 Water soluble
magnetic stirrer bar Silicone oil 0-250 Expensive
Glycerol 0-250 Water soluble
heating plate
variable speed control

the bath above 100°C, the water will boil and may spatter hot oil over
variable heating control you and the apparatus. Dispose of the contaminated oil into the
appropriate waste container, clean the bath with paper towels to absorb
the water and refill with fresh oil once completely dry.
• If the oil bath is a Pyrex@ dish, check the dish carefully for cracks and
'star' cracks because you do not do want the dish of hot oil to break.
• Support the stirrer hot plate on a 'labjack' so that you can quickly
remove the heat source if the reaction goes wrong (see mantles, p. 36).
• When the heating process is finished, allow the bath to cool and raise the
flask, let the oil drain from it into the bath and then wipe the flask with a
cloth or paper towel. Otherwise your hands, gloves, apparatus, com-
pounds, etc., may become contaminated with oil.
Electric heating mantles

These are used for heating round-bottom flasks only and come in various sizes.
Always used a heating mantle appropriate to the size of the flask you are going
to heat, since you need to control the heating process. The flask should fit snugly
into the mantle with the top half of the flask above the case of the mantle. If the
mantle is too small, heating will be inefficient, whereas if the mantle is too big,
the flask will be overheated and decomposition of the contents may occur.
Electric heating mantles comprise an electric heating element wrapped in a
glass fibre covering, protected by an earth screen and enclosed in an
aluminium or, more commonly, a polypropylene case to allow handling at
moderate temperatures. Heating control is provided either by a regulator
(b) built into the heating mantle (Fig. 5.13a) or by an external controller, which
is connected to the mantle by a plug.
Fig. 5.12 (a) A stirrer hot plate. (b) Using a
stirrer hot plate with an oil bath to heat a
Some mantles, 'stirrer mantles', have a built-in magnetic stirrer just like a
round-bottom flask. stirrer hot plate and can be used to mix the liquid using a magnetic flea or
bar (Fig. 5.13b). Stirrer mantles have two controls on the side: make sure that
you know the function of each, since one controls the extent of heating and
the other the stirrer speed.
When using mantles make the following safety checks:
• Make sure that the heating element is not damaged or worn. If in doubt
consult your instructor and get a replacement.
• Plug in and test the mantle controls ensuring that both heater and stirrer
are working. If fumes are given off from the heating elements someone
has spilt chemicals into the mantle: switch off, report the fault to your
instructor and obtain a replacement.
• Mantles are relatively slow to react to changes in the heater control
setting and it is easy to 'overshoot' the desired temperature. Therefore
always make small incremental changes in the heating control and if a
temperature below the boiling point of the liquid is required, make sure
that a thermometer is incorporated in the apparatus.

• When using a mantle with complex apparatus such as for distillation

(p. 107), support the mantle on a 'labjack' so that it can be removed
quickly if overheating occurs (Fig. 5.14).
• If the mantle is not equipped with a stirrer, remember to add anti-
bumping granules (p. 31) to the liquid.
heat control Hot-air guns

These can be used instead of Bunsen burners or micro burners provided that
the temperature required is not too high. The main uses of hot-air guns are
for drying glassware and as a heat source for distillation of liquids at
relatively moderate temperatures up to about 120°C.
Hot-air guns produce heated air, usually at two temperatures, and
cold air. As with a burner, the heat transferred to the flask being heated
can be controlled by 'waving' the hot air stream around the flask (see
heat control p. 32).
Remember that the hot-air gun has a hot-wire heating element; therefore
do not use it in the presence of flammable vapours. The nozzle of the hot-
stir control
air gun will become very hot and can cause burns or even ignite some
(bl solvents for some time after it has been switched off. You should switch the
gun to the cold-air mode for a few minutes after you have finished heating
5.13 Mantles: (a) heating mantle; (b) stirrer
Fig.
and place it in a 'holster' made from a support ring before finally turning
mantle.
off the gun.
Handling hot glassware

The safety precautions necessary for handling hot glassware depend upon:
• The temperature of the glassware.

• The type of glassware: test tube, beaker, conical flask, round-bottom
flask, etc.
• The size of the glassware.
• The manipulation being carried out.
Normally you do not need any hand protection to handle glassware at
temperatures up to 50 QC. For general-purpose work, such as removing
glassware from the drying oven, assembling hot glassware and manoeuvring
hot beakers and conical flasks to and from a burner, steam bath or hot plate,
heat-resistant gloves are suitable and should be available in the laboratory.
Where more intricate processes are required, such as hot filtration, then
mantle raised 'rubber fingers' made from medium-wall rubber tubing (Fig. 5.15) give
and lowered
adequate protection up to about 120 QC and are less cumbersome than
insulated gloves. 'Rubber fingers' are useful for small volumes, up to 150 mL,
of liquids when the flask or beaker can be easily held by one hand. If larger
volumes are being manipulated then two hands are required and heat-
resistant gloves are essential.
The following specific techniques should be noted:
adjustable
laboratory • Test tubes: should be held by a wooden test tube holder (Fig. 5.16), which
jack
provides adequate insulation and grip. You should never use folded pieces
of paper or metal tongs.
~ • Conical flasks: are often clamped to a support stand during heating and
to electrical supply
you should never attempt to use the clamp as a device to hold the flask
Fig. 5.14 Heating using a stirrer mantle. when removed from the support stand. If you place the flask on the
laboratory bench with the clamp attached it will fall over because of the
weight of the clamp. Furthermore, you will have little control over the

pouring process. Use 'rubber fingers' or an insulated glove and never use
folded pieces of paper or metal tongs (Fig. 5.17).
• Beakers: these have specific problems since they have no narrow neck
which can be gripped for lifting. Small beakers of volumes up to 400 mL
capacity can easily be gripped in one hand protected by an insulated
glove or 'rubber fingers'.
• Round-bottom flasks: small flasks of capacity up to 250 mL should be held
at the neck, gripped in one hand protected by an insulated glove or
'rubber fingers' and when moving and pouring from larger flasks they
should be held by the neck and supported underneath. Do not use a
clamp round the neck of the flask as a support.
Cooling
During laboratory work you will be required to carry out experiments at
Fig.5.15 Rubber 'fingers'.
temperatures below room temperature. The most common situations where
cooling is required are:
• Cooling solutions during recrystallization.

• Completion of precipitation in quantitatative (gravimetric) analysis and
preparations.
• Cooling exothermic reactions.
• Carrying out reactions at low temperatures.
There are three cooling media commonly used in the laboratory: crushed
ice, solid carbon dioxide (Dry Ice@, Drikold'P, Cardice'P) and liquid
nitrogen. You are unlikely to use liquid nitrogen in the undergraduate
laboratory - if it is required you must consult your demonstrator about the
special safety protocols required for its use.
The most suitable containers for cooling baths are plastic bowls (ice
baths), Pyrex@ dishes (solid CO2 baths) and Dewar flasks (solid CO2 and
liquid nitrogen baths). If Pyrex@ dishes are to be used below -20 QC, an
insulated container can be prepared by placing a smaller Pyrex@ dish
inside a larger one and filling the space between with an insulating
material such as cotton wool, cork chips or polyurethane foam chips.
Fig. 5.16 Holding a test tube. Remember that foam chips will dissolve if they come into contact with
many organic solvents.
If the temperature of the liquid or solution being cooled is critical,
do not assume that the temperature of the liquid or solution is the same
as that of the cooling bath: place a thermometer in the flask and
remember that for temperatures lower than -5 DC you should use an
alcohol thermometer (red thread) or a thermocouple-type thermometer
(after checking that the probe will not react with the contents of the
flask).
Ice baths
A slurry of crushed ice and water can be used to give a cooling bath in the
range 0 QC to 5 "C. Pure crushed ice does not give good contact with the
glassware and inefficient cooling results.
If temperatures below 0 QC are required, mixtures of crushed ice and
various inorganic salts can be used as shown in Table 5.2. Note that these
mixtures contain no liquid and therefore cooling is inefficient and the
temperatures indicated in the table are the lowest attainable under ideal
Fig.5.17 Pouring a hot solution. conditions.

Solid C02 baths

Table 5.2 Ice-salt mixtures Solid CO2, when mixed with organic solvents, provides cooling media of
temperatures ranging from -15°C to -100°C. Some common mixtures
together with the minimum achievable temperatures are shown in Table 5.3.
The CO2 and the organic solvent form a 'slush', which gives excellent contact
CaC12.6H20 1 : 2.5 -10°C with flasks and, therefore, efficient cooling.
NH4CI 1: 4 -15°C
NaCI 1:3 -20°C
CaCb.6H20 1 :0.8 -40°C Table 5.3 Solid COr solvent mixtures
Ethylene glycol -15°C Ice/NH4C1 cheaper

Acetonitrile -40°C Toxic, flammable
Chloroform -60°C Toxic
Ethanol -tz-c Flammable
Acetone -78°C Flammable
Diethyl ether -100°C Highly flammable
Solid CO2 is supplied as large hard blocks or small quantitres can be

prepared from cylinders of liquid CO2 as a 'snow'. Skin contact with solid
CO2 will cause frostbite and it must be handled with insulated gloves. The
cooling bath must be an insulated container, which will need to be topped up
with solid CO2 at regular intervals to maintain the temperature, or, if
prolonged cooling is required, a Dewar flask in which the coolant will
maintain its temperature for 12 hours or so.
To prepare a solid CO2 cooling bath:
• Choose the appropriate solvent and remember to take into account the
hazards associated with its use.
• Break the solid CO2 into small pieces. The CO2 'snow' can be broken
with a spatula, but the hard blocks should be wrapped in cloth and then
broken into pieces with a wooden or polyethylene mallet.
• Half fill the bath with the solvent and then, using an insulated glove, add
small pieces of the solid CO2 until the mixture stops 'boiling' and then
add a little more solid C02 and stir with a glass rod to give a slurry.
• Use an alcohol or thermocouple thermometer to check the temperature of
the bath.
• Top up the cooling bath with solid CO2 if the temperature begins to rise.
~ When using an internal thermometer to measure the

temperature of a liquid or solution which is being stirred with a mag-
netic flea or stirring bar, ensure that the thermometer bulb does not
come into contact with it.
To prevent the condensation of water into your flask, you should have an
inert gas flowing through it (see p. 126) and prepare the cold bath around the
flask and its contents so that slow cooling occurs. Sudden immersion of a
relatively 'hot' flask into the cold bath will cause the bath to 'boil' and air
(containing water vapour) will be sucked into the apparatus despite the inert
atmosphere. Alternatively a 'loosely packed' CaCb guard tube (see p. 117)
will suffice if cooling is not too rapid.
Cooling probes
These are rigid or flexible metal probes, which are connected to a
refrigeration compressor. The probe is placed in the cooling bath and covered
with a suitable solvent, which is then cooled to the temperature desired.

Cooling probes are commonly found in constant temperature baths, when a

temperature below ambient temperature is required, and probes can be used
instead of solid CO2 to produce temperatures down to -100 QC. Cooling
probes are expensive pieces of equipment; therefore you will find them
dedicated to a specific experiment and they are not usually available for basic
laboratory operations.
Drying
During your laboratory course it will be necessary to dry glassware, analytical
standard compounds (see p. 144), chemicals you have synthesized, crucibles
used in gravimetric analysis (see p. 139) and solvents. Drying solvents is
described on p. 127.
Drying glassware
For most general laboratory applications glassware can be dried in an electric
oven between 80 QC and 90 QC or by rinsing the glassware with a small
amount of water-miscible solvent, such as acetone or ethanol, and then
evaporating the solvent using a compressed-air jet. Remember to remove all
plastic components, taps and stoppers from the glassware before you put it in
the oven.
If glassware is required for anhydrous reactions, it must be heated in the
oven above 100 QC, assembled while hot and allowed to cool while a stream
of inert gas is passed through it (see p. 126).
Drying solids
Here the term 'drying' means removal of a solvent, not specifically water,
from a solid by evaporation. The rate of evaporation and thus the rate of
drying can be increased by one (or all) of the following:
• Heating the chemical.

• Using a drying agent in a closed container to absorb the solvent.
• Reducing the atmospheric pressure.
Only chemicals which are thermally stable should be dried by heating.
Most inorganic compounds, which are salts with relatively high melting
points, can be dried in an electric oven to remove water, whereas organic
compounds, many of which have relatively low melting points, need to be
treated with more care and the oven temperature should be set between 30 QC
and 50 QCbelow the melting point of the chemical. Chemicals must be placed
glass in the oven on a clock-glass or watch-glass and be spread as thinly as
perforated plate possible, to increase the rate of solvent evaporation.
or gauze If you cannot dry your compound in the oven, then use a desiccator.
desiccant Desiccators are made from glass or plastic and some, vacuum desiccators, are
equipped with a tap to allow evacuation as shown in Fig. 5.18. The bottom
of the desiccator is filled with a drying agent (desiccant) and the chemical, on
a watch-glass or clock-glass, is placed on the mesh shelf above and the
plastic desiccator closed by sliding the lid onto the desiccator to provide an air-tight
seal. The desiccant absorbs the solvent from the 'atmosphere' in the
desiccator as it evaporates from the solid. The nature of the desiccant
depends upon the solvent to be removed (Table 5.4).
The most common drying agents for removal of water are anhydrous
CaCb and self-indicating silica gel. The CaCb should have the appearance of
Fig. 5.18 Desiccators. 'chalky' lumps. Self-indicating silica gel is blue in the 'active' state and pink

when its capacity for water absorption is 'exhausted'. The water can be
removed from the pink silica gel by heating in an oven above 100°C and
restoration of the blue colour indicates reactivation.
Table 5.4 Drying agents for desiccators
Silica gel
CaCI2
solid KOH Corrosive
P20S Corrosive
Cone. H2S04 Corrosive liquid
EtOH, MeOH CaCI2
Hydrocarbons Paraffin wax
The rate of drying can be increased by evacuating the desiccator and

vacuum desiccators are specially designed for this purpose. The principal
steps for the use of a vacuum desiccator are as follows and it is essential that
you follow the order of the operations:
• Check that the appropriate desiccant is present and active.

• Check that the desiccator seals perfectly: ensure that the ground-glass
edges to the lid are greased lightly or, if the desiccator has a rubber
gasket, carry out a trial evacuation to ensure that the vacuum seals the
desiccator by gently pressing the lid onto the gasket - listen for air being
sucked around the seal.
• Place the sample onto a watch- or clock-glass or a beaker covered with
tissue paper (secure with an elastic band) and place it on the shelf.
• Place the lid on the desiccator and open the tap and cover the desiccator
with an appropriately sized safety cage (Fig. 5.19) to prevent injury from
flying glass in the case of an implosion.
vacuum
stopcock
hole for
to vacuum
stopcock
Fig. 5.19 Vacuum desiccator (with mesh safety cage).
• Connect the tap to an operating source of vacuum: a water pump (p. 30)
or vacuum pump (p. 110) and open the tap slowly to evacuate the
desiccator.
• When drying appears complete, close the tap on the dessicator and then
disconnect the vacuum supply.

• Place a small piece of filter paper on the end of the tap and open the tap
slowly. The filter paper will stick to the tap as air is drawn slowly
through it. When the air has completely filled the desiccator, the filter
paper will fall off and it is safe to open the desiccator.
If you need to heat the compound under vacuum, then you will need to use a
vacuum oven (for large quantities of solids). The principles of operation of
these pieces of equipment are similar to those of a vacuum desiccator, except
that an electric heater is incorporated. Always allow the apparatus to cool to
room temperature before releasing the vacuum.
Drying liquids
This usually means removing water from a liquid chemical or a solution of a
chemical in a water-immiscible solvent. You will always need to dry solutions
after a liquid-liquid extraction (p. 102) and you may need to dry liquids after
evaporation (p. 121) or distillation (p. 107). In both cases the liquid is placed in
direct contact with the solid drying agent, i.e. the drying agent is added to the
liquid or solution. Ideally the drying agent should be totally insoluble in the
liquid, should not react with it, absorb the water quickly and efficiently, and be
easily filtered off. A list of the common drying agents is given in Table 5.5.
Table 5.5 Drying agents for liquids and solutions
MgS04 High Fast Good Best general use

Na2S04 High Slow Poor Useful
CaCI2 High Slow Poor Reacts with 0 and N compounds
CaS04 Low Fast Good Useful
K2C03 High Fast Good Reacts with acidic compounds
Molecular sieve Moderate Fast Good Must be activated at 300 DC
Capacity: amount of water taken up.

Speed: rate of water absorption.
Efficiency: extent of drying after treatment.
You must remember that the drying agent will absorb some of the liquid
or solvent being dried as well as the water. If you wish to dry a small volume
of liquid, it is better to dissolve it in a low-boiling water-immiscible solvent
and dry the solution by the procedure described in Box 5.3.

Jointed glassware
This type of glassware, commonly known as Quickfit'[', comprises a complete
range of components fitted with standard-taper ground-glass joints. The
joints are fully interchangeable with those of the same size and apparatus for
a whole range of experiments can be assembled from the simple components
without the need to use rubber bungs, corks, etc. Where there is a mismatch
between the sizes of the joints of the pieces of glassware, reduction and
expansion adapters can be used. A typical range of jointed glassware is
illustrated in Fig. 5.20.
06round-bottom flasks three-neck round-bottom flask
addition funnel seperatorv funnel
air condenser
(or a fractionation column)
guard tube stopper reduction/expension adapters
still-head splash-head Claisen adapter
distillation vacuum gas inlet thermometer

adapter distillation adapter adapter
adapter
Fig. 5.20 Glass equipment with standard-taper ground-glass joints.

Basic laboratory procedures I1
The ground-glass joint on the glassware is classified according to the

diameter of the joint at its widest point (internal diameter) and the length of
the ground-glass portion of the joint. Thus a 14/23 joint has a maximum
internal diameter of 14mm and a length of 23 mm. Other common joint sizes
you will frequently encounter are 19/26, 24/29 and 35/39. The joint size is
always etched into glass on the side of or near to the joint. For obvious
reasons, joints are categorized as 'female' and 'male'.
Care of jointed glassware

Jointed glassware is much more expensive than ordinary glassware because of
the precision required in fabricating the joints. If the joints 'seize' and cannot
be separated the glassware cannot be used again and you may have the
problem of a stoppered flask containing a volatile organic solvent, which
somebody has to open! If this happens, consult your instructor for help and
further advice.
There are two main causes of 'seized' joints:
1. Using solutions of potassium hydroxide or sodium hydroxide in water or
other solvents, which attack the glass.
2. Trapping chemicals, including solids and solutions of solids, in the
ground-glass joints.
If you are using jointed glassware with strong alkalis (NaOH, KOH), you
must grease the joints. In most cases a simple hydrocarbon-based grease, such
as petroleum jelly, will suffice, since it is easily removed from the joints by
wiping with a cloth wet with a hydrocarbon solvent (petroleum spirit, b.pt. 60-
80 QC).Avoid silicone-based grease, since this is difficult to remove, soluble in
some organic solvents and may contaminate your reaction products.
To grease a joint, put a small smear of grease on the upper part of the
'male' joint, push it into the 'female' joint with a twisting movement and the
joint should become 'clear' from the top to about half-way down. If more
than half the joint has become 'clear', you have used too much grease:
separate the joints, clean with a solvent-soaked cloth and repeat the process.
To avoid trapping chemicals in the ground-glass joints, fill flasks etc. using
a long-stemmed filter funnel or paper cone, which extends past the joint into
the flask (see p. 25).
Screwcap adapters
Screwcap or thermometer adapters allow you to place thermometers, glass
tubes or air bleeds into jointed glassware flasks. The screwcap adapter works
by using the screwcap to compress a rubber ring round the thermometer or
glass tube and thus hold it in place. The flexibility of the system allows the
height of the thermometer/glass tube to be varied. The adapters come in
different joint sizes and varying hole sizes to accommodate different-diameter
thermometers, tubes, Pasteur pipettes, etc. The component parts of a
screwcap adapter are shown in Fig. 5.21.
To use the screwcap adapter with a thermometer:
• Always disassemble the adapter to ensure that the rubber ring and the
Teflon® seal are present. If they are missing, get replacements before use.
The Teflon® seal is to protect the rubber ring from corrosive and solvent
vapours.
• Ease the rubber ring onto the thermometer (see p. 13) and slide the
Teflon'f seal on below the ring.

• Slide the screwcap over the top of the thermometer and then screw the
screwcap
whole assembly onto the base of the adapter and tighten the screw
slightly, just enough to hold the thermometer.
~ rubber ring
• Trial fit the adapter and thermometer into the apparatus and adjust the
~ ( Teflon® seal
height of the thermometer by disassembling the adapter and moving the
rubber ring up or down. Reassemble the adapter.
• Check for final fit and tighten the screwcap firmly, but do not over-tighten.
Joint clips
Plastic joint clips or Keck@ clips (Fig. 5.22a) are used for holding ground-
joint glass joints firmly together and may be used to replace clamps and support
stands at certain points when building apparatus (see p. 109) and are essential
when using rotary evaporators (p. 122). The main weakness of these
Fig. 5.21 The screwcap adapter. otherwise useful devices is that they soften at about 130 QC and this may
allow the joints to separate. Therefore they should never be used at the 'hot
end' of a distillation, for example. The clip should be used to hold a
distillation adapter on the end of a water condenser, or the flasks onto a
fraction collector, but never on the distilling flask or to hold the condenser
onto the still head (Fig. 5.22b).
When using joint clips always:
• Check that you are using the appropriate size of clip for the joints being
held. The clips are often colour coded.
• Check that the clip is not cracked or split.
• Check that the wide 'lower jaw' of the clip fits under the rim of the
'female' joint and the 'upper jaw' fits round the male joint.
• Support the joints with your hand as you push the clip into place. If in
doubt use a protective insulated glove.
rrrr
joint clips here
in
(b)
Fig. 5.22 Joint clips and their use.

-------8 Principles of solution chemistry
A solution is a homogeneous liquid, formed by the addition of solutes to a

solvent. The behaviour of solutions is determined by the type of solutes
involved and by their proportions, relative to the solvent. Many laboratory
exercises involve calculation of concentrations, e.g. when preparing an
experimental solution at a particular concentration, or when expressing data
in terms of solute concentration. Make sure that you understand the basic
principles set out in this chapter before you tackle such exercises.
Solutes can affect the properties of solutions in several ways, as follows.
Electrolytic dissociation
This occurs where a substance dissociates to give charged particles (ions). For
a strong electrolyte, e.g. Na+CI-, dissociation is essentially complete. In
contrast, a weak electrolyte, e.g. ethanoic acid, will be only partly dissociated,
depending upon the pH and temperature of the solution (p. 57).
Osmotic effects
These are the result of solute particles lowering the effective concentration of
the solvent (water). These effects are particularly relevant to biological
systems since membranes are far more permeable to water than to most
solutes. Water moves across biological membranes from the solution with the
higher effective water concentration to that with the lower effective water
concentration (osmosis).
Ideal/non-ideal behaviour
This occurs because solutions of real substances do not necessarily conform
to the theoretical relationships predicted for dilute solutions of so-called ideal
solutes. It is often necessary to take account of the non-ideal behaviour of
real solutions, especially at high solute concentrations (see Lide (2000) for
appropriate data).
Concentration
In SI units (p. 71) the concentration of a solute is expressed in mol m -3,
which is essential for calculating specific parameters for substances (e.g.
p. 73), but which is inconvenient when dealing with solutions in the labora-
tory. A cubic metre (m") of water weighs approximately I ton! A common
unit of volume used in chemistry is the litre (L): this is a non-SI unit and is
converted to the SI unit of volume (m") using 1.0 L = 10-3 m". The concen-
tration of a solute is usually symbolized by square brackets, e.g. [NaCI].
Details of how to prepare solutions are given on pp. 17, 19.
A number of alternative ways of expressing the relative amounts of solute
and solvent are in general use, and you may come across these terms in your
practical work or in the literature.
Molarity
This is the term used to denote molar concentration, [C), expressed as moles
of solute per litre volume of solution (mol L -1). This non-SI term continues
to find widespread usage, in part because of the familiarity of working
scientists with the term, but also because laboratory glassware is calibrated in

Principles of solution chemistry
millilitres and litres, making the preparation of molar and millimolar

solutions relatively straightforward. However, the symbols in common use
for molar (M) and millimolar (mM) solutions are at odds with the SI system
and many people now prefer to use mol L -I and mmol L -I respectively, to
avoid confusion. Box 6.1 gives details of some useful approaches to calcula-
tions involving molarities.
Molality
This is used to express the concentration of solute relative to the mass of solvent,
i.e. mol kg:". Molality is a temperature-independent means of expressing solute
concentration, rarely used except when the osmotic properties of a solution are
of interest (p. 49).
Per cent composition (% wlw)

This is the solute mass (in g) per 100 g solution. The advantage of this
expression is the ease with which a solution can be prepared, since it simply
requires each component to be pre-weighed (for water, a volumetric
measurement may be used, e.g. using a measuring cylinder) and then mixed
together. Similar terms are parts per thousand (%0), i.e. mg g", and parts per
million (ppm), i.e. J.lgg-I.

Per cent concentration (% w]v and % vlv )

For solutes added in solid form, this is the number of grams of solute
per 100 mL solution. This is more commonly used than per cent com-
position, since solutions can be accurately prepared by weighing out the
required amount of solute and then making this up to a known volume
using a volumetric flask. The equivalent expression for liquid solutes is
% v/v.
The principal use of mass/mass or mass/volume terms (including g L -1)
is for solutes whose molecular mass is unknown (e.g. polymers), or for
mixtures of certain classes of substance (e.g. total salt in sea water). You
should never use the per cent term without specifying how the solution
was prepared, i.e. by using the qualifier w/w, w]» or v]v, For mass
concentrations, it is often simpler to use mass per unit volume, e.g.
mgL -I, jJ.gjJ.L-1, etc.
Parts per million concentration (ppm)

This is a non-SI weight per volume (w/v) concentration term commonly used
in quantitative analysis such as flame photometry, atomic absorption
spectroscopy and gas chromatography, where low concentrations of solutes
are to be analysed. The term ppm is equivalent to the expression of
concentration as IlgmL-1 (l0-6gmL-1) and a 1.0ppm solution of a
substance will have a concentration of 1.0jJ.gmL-1 (1.0 x 1O-6gmL-1). A
typical procedure for calculations in terms of ppm is shown in Box 6.2.
Parts per billion (ppb) is an extension of this concentration term as
ng mL -I (10-9 g mL -1) and is commonly used to express concentrations
of very dilute solutions. For example, the allowable concentration of

arsenic in water may be 0.05 ppm, but it is more conveniently expressed

as 50 ppb.
Activity (a)
This is a term used to describe the effective concentration of a solute. In
dilute solutions, solutes can be considered to behave according to ideal
(thermodynamic) principles, i.e. they will have an effective concentration
equivalent to the actual concentration. However, in concentrated solutions
( ~ 0.5 mol L -I), the behaviour of solutes is often non-ideal, and their
effective concentration (activity) will be less than the actual concentration [C].
Table 6.1 Activity coefficient of NaCI solutions The ratio between the effective concentration and the actual concentration is
as a function of molality. Data from Robinson called the activity coefficient (I') where
and Stokes (1970)
a
I' = [e] [6.3]
0.1 0.778 Equation [6.3] can be used for SI units (mol m "), molarity (mol L -1) or
0.5 0.681 molality (mol kgr '). In all cases, l' is a dimensionless term, since a and [C] are
1.0 0.657 expressed in the same units. The activity coefficient of a solute is effectively
2.0 0.668
4.0 0.783 unity in dilute solution, decreasing as the solute concentration increases
6.0 0.986 (Table 6.1). At high concentrations of certain ionic solutes, I' may increase to
become greater than unity.
~ Activity is often the correct expression for theoretical

relationships involving solute concentration (e.g. where a property of
the solution is dependent on concentration). However, for most practical
purposes, it is possible to use the actual concentration of a solute rather
than the activity, since the difference between the two terms can be ig-
nored for dilute solutions.
Equivalent mass (equivalent weight)

Equivalence and normality are outdated terms, although you may come
across them in older texts. The magnitude of an equivalent mass (equivalent
weight) can be simply identified from the balanced equation for the reaction
being considered. Remember that the equivalent mass can change, depending
on the reaction, as the following reactions illustrate.
For:
HCI + NaOH ---+ NaCI + H20
1mol of HCI reacts with 1mol of NaOH, the equivalent mass of HCI is
M,= 36.5 and the equivalent mass of NaOH is also its M; = 40.
For:
H2S04 + 2NaOH ---+ Na2S04 + 2H20
since 1mol of H2S04 reacts with 2 mol of NaOH, the equivalent mass of
H2S04 is M, 7 2 = 98 7 2 = 49, while the equivalent mass of NaOH is still
M, = 40.
For:
5FeS04 + KMn04 ---+ Fe2(S04)3 + 2MnS04
since 1mol of KMn04 reacts with 5 mol of FeS04, then the equivalent mass
ofKMn04 is M, 7 5 = 15875 = 31.6, and that of FeS04 is still M, = 152.
But, for:
H2S04 + Na2C03 ---+ Na2S04 + CO2 + H20
since the reaction is 1:1, the equivalent masses of H2S04 and Na2C03 are
their Mr values, 98 and 106 respectively.

As a result of this possible confusion, the concept of equivalent mass

(weight) is rarely used.
Normality
A 1 normal solution (1 N) is one that contains one equivalent mass of a
substance per litre of solution. The general formula is:
, mass of substance per litre
norma Iity = ----, ------- [6.4]
equivalent mass
Osmolarity
This non-SI expression is used to describe the number of moles of
osmotically active solute particles per litre of solution (osmol L:"). The need
for such a term arises because some molecules dissociate to give more than
one osmotically active particle in aqueous solution.
Osmolality
This term describes the number of moles of osmotically active solute particles
per unit mass of solvent (osmol kg "). For an ideal solute, the osmolality can
be determined by multiplying the molality by n, the number of solute
particles produced in solution (e.g. for NaCl, n = 2). However, for real
solutes, a correction factor (the osmotic coefficient, 1J) is used:
osmolality = molality x n x 1J [6.5]
Table 6.2 Osmotic coefficients of NaCI If necessary, the osmotic coefficients of a particular solute can be obtained
solutions as a function of molality, Data from from tables (e.g. Table 6.2): non-ideal behaviour means that 1J may have
Robinson and Stokes (1970) values> 1 at high concentrations. Alternatively, the osmolality of a solution
can be measured using an osmometer.
0.1 0.932 Osmotic pressure

0.5 0.921
1.0 0.936
This is based on the concept of a membrane permeable to water, but not to
2.0 0.983 solute molecules. For example, if a sucrose solution is placed on one side and
4,0 1.116 pure water on the other, then a passive driving force will be created and
6.0 1.271
water will diffuse across the membrane into the sucrose solution, since the
effective water concentration in the sucrose solution will be lower. The
tendency for water to diffuse into the sucrose solution could be counteracted
by applying a hydrostatic pressure equivalent to the passive driving force.
Thus, the osmotic pressure of a solution is the excess hydrostatic pressure
required to prevent the net flow of water into a vessel containing the
solution. The SI unit of osmotic pressure is the pascal, Pa (= kg m-I s-2).
Older sources may use atmospheres, or bars, and conversion factors are given
in Box 9.1 (p. 72). Osmotic pressure and osmolality can be interconverted
using the expression 1 osmol kg-I = 2.479MPa at 25°C.
The use of osmotic pressure has been criticized as misleading, since a
solution does not exhibit an 'osmotic pressure' unless it is placed on the other
side of a selectively permeable membrane from pure water!
Colligative properties and their use in osmometry

Several properties vary in direct proportion to the effective number of
osmotically active solute particles per unit mass of solvent and can be used to
determine the osmolality of a solution. These colligative properties include
freezing point, boiling point and vapour pressure.

E 4
~
'"Q;
E
0-
E
r Fig. 6.7 Temperature responses of a

cryoscopic osmometer. The response
can be subdivided into:
1. initial supercooling
2. initiation of crystallization
3. crystallization/freezing
4. plateau, at the freezing point
time 5. slow temperature decrease
An osmometer is an instrument which measures the osmolality of a

solution, usually by determining the freezing point depression of the solution
in relation to pure water, a technique known as cryoscopic osmometry. A
small amount of sample is cooled rapidly and then brought to the freezing
point (Fig. 6.1), which is measured by a temperature-sensitive thermistor
probe calibrated in mosmol kg ". An alternative method is used in vapour
pressure osmometry, which measures the relative decrease in the vapour
pressure produced in the gas phase when a small sample of the solution is
equilibrated within a chamber.
Solubility
The extent to which a solute will dissolve in a solvent is called its solubility.
The solubility of a chemical is conventionally expressed as the maximum
number of grams of a chemical that will dissolve in lOOg of solvent but
conversion to mol L -I or g L -I is simple and may be appropriate for some
applications (see below). Since solubility is temperature dependent, is always
quoted at a specific temperature. With a very few exceptions, increasing the
temperature of a solvent increases the solubility of the solute.
Saturated solutions
For practical purposes, a saturated solution is one in which no more solute will
dissolve. For example, the solubility of sodium chloride in water is 35.6 g per
lOOg at 25 QC and 39.1 g per 100 g at 100 QCand both solutions are saturated
solutions at their respective temperatures. If the 100 QC solution is cooled to
25 QC, then 3.5 g of NaCl crystals will precipitate from the solution, because
the solution at 25 QCrequires only 35.6 g of NaCI for saturation. This process
is the basis of purification of compounds by recrystallization (see p. 92).
Solubility product
In dilute aqueous solutions, it has been demonstrated experimentally for
poorly soluble ionic salts (solubilities less than 0.01 mol L -I) that the
mathematical product of the total molar concentrations of the component
ions is a constant at constant temperature. This product, K; is called the
solubility product. Thus for a saturated solution of a simple ionic compound
AB in water, we have the dynamic equilibrium:

ABsolid====; Atq) + B~q)

where AB represents the solid which has not dissolved, in equilibrium with its
ions in the aqueous saturated solution. Then:
For example, silver chloride is a solid of solubility 0.000 15g per 100mL of
water in equilibrium with silver cations and chloride ions. Then:
K; = [Ag+] x [Cl"]
The solubility of AgCl is 0.0015gmL-1 (l0 x solubility per 100g, assuming

that the density of water is 1.0g mL -I) and therefore the solubility of AgCl is
0.0015 -i- 143.5 = 1.05 x 10-5 mol L -I. Thus the saturated solution contains
1.05 x 10-5 mol L -1 of Ag" ions and 1.05 x 10-5 mol L -1 of Cl- ions and
the solubility product Ks is
Ks = (1.05 x 10-5) x (1.05 x 10-5) = 1.1 X 10-10 mol? L-2
If the solid does not have a simple I: I ratio of its ionic components, e.g.
PbCb, then the solubility product is given by:
K; = [Pb2+] x [Cn2
In general terms, the solubility product for a compound MyNx, is given by

x, = [M+Y x [N-Y
The practical effects of solubility products are demonstrated in the

detection of anions and cations by precipitation (p. 135) and in quantitative
gravimetric analysis (p. 139). For example, if dilute aqueous solutions of
silver nitrate (solubility 55.6 g per 100g of water) and sodium chloride
(solubility 35.6 g per 100g of water) are mixed, an immediate white
precipitate of AgCl is produced because the solubility product of AgCI has
been exceeded by the numbers of Ag" and CI- ions in the solution, even
though the ions come from different 'molecules'. A saturated solution of
AgCl is formed and the excess AgCI precipitates out. The solubility product
of the other combinations of ions is not exceeded and thus sodium and
nitrate ions remain in solution. Even if the concentration of Ag" is extremely
low, the solubility product for AgCl can be exceeded by the addition of an
excess of Cl- ions, since it is the multiplication of these two concentrations
which defines the solubility product. Thus soluble chlorides can be used to
detect the presence of Ag" ions and, conversely, soluble silver salts can be
used to detect Cl- ions, both quantitatively and qualitatively.
Reactions of ions in solution

There are essentially only four basic reactions of ions in solution:
1. Acid-base reactions.
2. Precipitation reactions.
3. Complexation reactions
4. Reduction-oxidation (redox) reactions.
Acid-base, precipitation and complexation reactions are all examples of

exchange (metathesis) reactions in which ions in solution 'exchange partners',
for example:
A+Y- +B+Z- --> A+Z- +B+Y-

Metathesis reactions are really equilibria between the ionic species, which
are displaced to the right (to the reaction product) by a feature which defines
the classification of the reaction type.
Acid-base reactions
The most common acid-base reactions are exemplified by the neutralization
reaction between hydrogen ions and hydroxide ions; for example, the
reaction between dilute hydrochloric acid and dilute sodium hydroxide:
H+CI(;.q)+ Na+OH(;.q) -+ H-OH(liq) + Na+CI(;.q)
Since water is essentially a covalent compound (see p. 56) its formation
effectively removes H+ and OH- from the equilibrium and drives the reaction
to completion. Other examples of this general type of reaction include the
removal of a molecule as a gas, such as reactions of acids with carbonates
and bicarbonates, where unstable H2C03 decomposes to H20 and CO2.
Precipitation reactions
In these reactions between ions, one substance is removed from the ionic
equilibrium by precipitation (see solubility product, p. 50) and drives the
equilibrium to the right (see p. 51).
Complexation reactions
A complex ion is formed by the reaction of a metal cation, in particular
transition metals, with an electron donor molecule (ligand), which can be
neutral or have a negative charge. The cation can accept an electron pair and
the ligand donates an electron pair to form a covalent donor (co-ordinate)
bond between the ligand and the metal ion. The ligands are said to co-
ordinate with the metal ion to give a complex. Many ligands are more
powerful electron donors than water and thus the addition of a ligand to an
aqueous solution of a metal cation displaces the equilibrium towards the
more stable complex ion. (See p. 151 for stability constants and complexes.)
The effects of complex formation are illustrated in Box 6.3.
The overall effect of complex formation is to 'remove' a hydrated metal
ion from the mixture of ions in solution by displacing the equilibrium in
favour of the complex, cf. the similar process in the formation of water in
acid-base titrations and precipitation reactions.
Reduction-oxidation (redox) reactions

The concepts of oxidation and reduction are defined in terms of complete
electron transfer from one atom, ion or molecule of a chemical to another:
• Chemical oxidized - chemical loses electron(s).
• Chemical reduced - chemical gains electron(s).
This approach is generally applicable to most reactions and avoids compli-
cations of the older definitions involving hydrogen and oxygen. You should
realize that if a chemical is oxidized during a reaction, then another must be
reduced: oxidation and reduction always occur together. Furthermore:
• Oxidizing agent - gains electron(s) and is therefore reduced.
• Reducing agent -loses electron(s) and is therefore oxidized.
The following reaction between magnesium metal and dilute acid illustrates
these concepts:

Magnesium metal has lost two electrons in forming Mg2+ ions and has
therefore been oxidized. The two protons have each gained an electron to
form hydrogen atoms (and then one hydrogen molecule) and have been
reduced. Since magnesium metal has been oxidized, it is a reducing agent and
because H+ has gained an electron, it is an oxidizing agent.
The stoichiometry of a redox reaction is defined by the number of
electrons transferred between the oxidizing agent and the reducing agent since
the number of electrons lost by the reducing agent must equal the number of
electrons gained by the reducing agent, e.g.
2Mg(s) + 02(g) --> 2MgO(s)

So that you can work out titrations involving redox reactions, you will find
it necessary to balance redox equations, and while it is easy for simple
reactions such as those above, more complex redox reactions, such as the one
below, require more thought and work.
Such problems can be broken down into several simple steps, each with its
own set of rules:
• Identify the atoms, IOns or molecules which have been oxidized and
reduced.
• Identify the ionic half-reactions for the species being oxidized and
reduced and combine them.
• Balance the ionic half-reactions and combine them to give a balanced
equation for the reaction.
The species which are oxidized and reduced can be identified using the
concept of oxidation numbers. The rules for determining oxidation numbers
and examples are given in Box 6.4 and the application of ionic half-reactions

to balance redox equations is shown in Box 6.5. Note that the result of the
use of partial ionic equations gives a balanced ionic equation for the redox
reaction.
~ In simple acid-base, precipitation and complexation

reactions, no change of oxidation number occurs at any of the atoms
involved.


-------8 pH and buffer solutions
pH is a measure of the amount of hydrogen ions (H+) in a solution. It is

usual to think of aqueous solutions as containing H+ ions (protons), though
protons actually exist in their hydrated form, as hydronium ions (H30+). The
proton concentration of an aqueous solution [H+] is affected by several
factors:
• Ionization (dissociation) of water, which liberates protons and hydroxyl

ions in equal quantities, according to the reversible relationship:
[7.1]
• Dissociation of acids, according to the equation:
H-A ~ H+ +A- [7.2]
where H-A represents the acid and A-is the corresponding conjugate
base. The dissociation of an acid in water will increase the amount of
protons, reducing the amount of hydroxyl ions as water molecules are
formed (eqn [7.1]). The addition of a base (usually, as its salt) to water
will decrease the amount of H+, owing to the formation of the conjugate
acid (eqn [7.2]).
• Dissociation of alkalis, according to the relationship:
[7.3]
where X+OH- represents the un dissociated alkali. Since the dissociation

of water is reversible (eqn [7.1]), in an aqueous solution the production of
hydroxyl ions will effectively act to 'mop up' protons, lowering the
proton concentration.
Many compounds act as acids, bases or alkalis: those which are almost
completely ionized in solution are usually called strong acids or bases, while
weak acids or bases are only slightly ionized in solution (p. 45).
In an aqueous solution, most of the water molecules are not ionized. In
fact, the extent of ionization of pure water is constant at any given
temperature and is usually expressed in terms of the ion product (or
ionization constant) of water, Kw:
[7.4]
where [H+] and [OH-] represent the molar concentration (strictly, the
activity) of protons and hydroxyl ions in solution, expressed as mol L -I. At
25 QC,the ion product of pure water is 10-14 mol/ L -2 (i.e. 10-8 mof mr").
This means that the concentration of protons in solution will be 10-7 mol
L -1 (10-4 mol m-3), with an equivalent concentration of hydroxyl ions (eqn
[7.1]). Since these values are very low and involve negative powers of 10, it is
customary to use the pH scale, where:
[7.5]
and [H+] is the proton activity (see p. 48).

pH and buffer solutions
Table 7.1 Effects of temperature on the ion ~ While pH is strictly the negative logarithm (to the
product of water (Kw>' H+ ion concentration base 10) of H+ activity, in practice H+ concentration in mol l,r ' (equiva-
and pH at neutrality. Values calculated from lent to kmol m-3 in SI terminology) is most often used in place of activ-
Lide (2000). ity, since the two are virtually the same, given the limited dissociation of
H20. The pH scale is not SI: nevertheless, it continues to be used widely
in chemistry.
The value where an equal amount of H+ and OH- ions are present is
0 0.11 X 10~4 33.9 7.47 termed neutrality: at 2S QCthe pH of pure water at neutrality is 7.0. At this
4 0.17 x 10~4 40.7 7.39
0.29 x 10~4
temperature, pH values below 7.0 are acidic while values above 7.0 are
10 53.7 7.27
20 0.68 x 10~4 83.2 7.08 alkaline. However, the pH of a neutral solution changes with temperature
25 1.01 x 10~4 100.4 7.00 (Table 7.1), owing to the enhanced dissociation of water with increasing
30 1.47 X 10~4 120.2 6.92
37 2.39 x 10-4
temperature. This must be taken into account when measuring the pH of any
154.9 6.81
45 4.02 x 10-4 199.5 6.70 solution and when interpreting your results.
Always remember that the pH scale is a logarithmic one, not a linear one:
a solution with a pH of 3.0 is not twice as acidic as a solution of pH 6.0, but
a thousand times as acidic (i.e. contains 1000 times the amount of H+ ions).
Therefore, you may need to convert pH values into proton concentrations
before you carry out mathematical manipulations (see Box 40.2). For similar
reasons, it is important that pH change is expressed in terms of the original
and final pH values, rather than simply quoting the difference between the
values: a pH change of 0.1 has little meaning unless the initial or final pH is
known.
Measuring pH
pH electrodes
Accurate pH measurements can be made using a pH electrode, coupled to
Table 7.2 Properties of some pH indicator a pH meter. The pH electrode is usually a combination electrode,
dyes comprising two separate systems: an H+ -sensitive glass electrode and a
reference electrode which is unaffected by H+ ion concentration (see Fig.
7.2). When this is immersed in a solution, a pH-dependent voltage
between the two electrodes can be measured using a potentiometer. In
most cases, the pH electrode assembly (containing the glass and reference
Thymol blue
(acid) Red-yellow 1.2-6.8 electrodes) is connected to a separate pH meter by a cable, although some
Bromophenol hand-held instruments (pH probes) have combination electrodes and
blue Yellow-blue 1.2-6.8 meter within the same assembly, often using an H+ -sensitive field effect
Methyl orange Red-yellow 2.8-4.0 transistor in place of a glass electrode, to improve durability and port-
Congo red Blue-red 3.0-5.2 ability.
Bromocresol Box 7.1 gives details of the steps involved in making a pH measurement
green Yellow-blue 3.8-5.4 with a glass pH electrode and meter.
Methyl red Red-yellow 4.3-6.1
Litmus Red-blue 4.5-8.3 pH indicator dyes
Chlorophenol red Yellow-red 4.8-6.3 These compounds (usually weak acids) change colour in a pH-dependent
Bromocresol manner. They may be added in small amounts to a solution, or they can be
purple Yellow-purple 5.2-6.8
used in paper strip form. Each indicator dye usually changes colour over a
Bromothymol
blue Yellow-blue 6.0-7.6 restricted pH range (Table 7.2): universal indicator dyes/papers make use of a
Neutral red Red-yellow 6.8-8.0 combination of individual dyes to measure a wider pH range. Dyes are not
Phenol red Yellow-red 6.8-8.2 suitable for accurate pH measurement as they are affected by other
1-Naphthol- components of the solution including oxidizing and reducing agents and salts.
phthalein Yellow-blue 7.2-8.6 However, they are useful for:
Phenol-
phthalein None-red 8.3-10.0 • estimating the approximate pH of a solution;
• determining a change in pH, e.g. at the end-point of a titration.

Buffers
Rather than simply measuring the pH of a solution, you may wish to control
the pH, during EDTA complexation titrations (see p. 152) or preparative
experiments involving carbonyl compounds, and one of the most effective
ways to control pH is to use a buffer solution.
A buffer solution is usually a mixture of a weak acid and its conjugate
base. Added protons will be neutralized by the anionic base while a reduction
in protons, e.g. due to the addition of hydroxyl ions, will be counterbalanced
by dissociation of the acid (eqn [7.2]); thus the conjugate pair acts as a
'buffer' to pH change.
The British standard for the pH scale is an aqueous solution of potassium
hydrogen phthalate (0.05 M), which has a pH of 4.001 at 20 QC and is often
used as a calibration solution for pH meters.
Buffer capacity and the effects of pH

The extent of resistance to pH change is called the buffer capacity of a
solution. The buffer capacity is measured experimentally at a particular pH
by titration against a strong acid or alkali: the resultant curve will be strongly
sigmoidal, with a plateau where the buffer capacity is greatest (Fig. 7.1). The
mid-point of the plateau represents the pH where equal quantities of acid and
conjugate base are present, and is given the symbol pKa, which refers to the
negative logarithm (to the base 10) of the acid dissociation constant, Ka,
where
alkali added
[7.6]
Fig. 7.1 Theoretical pH titration curve for a
buffer solution. pH change is lowest and buffer By rearranging eqn [7.6] and taking negative logarithms, we obtain:
capacity is greatest at the pKa of the buffer
solution. [A-]
pH = pKa + 10glO [HA] [7.7]
This relationship is known as the Henderson-Hasselbalch equation and it

shows that the pH will be equal to the pKa when the ratio of conjugate base
to acid is unity, since the final term will be zero. Consequently, the pKa of a
buffer solution is an important factor in determining the buffer capacity at a
particular pH. In practical terms, this means that a buffer solution will work
most effectively at pH values about one unit either side of the pKa.
Selecting an appropriate buffer

When selecting a buffer, you should be aware of certain limitations to its use.
Citric acid and phosphate buffers readily form insoluble complexes with
divalent cations, while phosphate can also act as a substrate, activator or
inhibitor of certain enzymes. Both of these buffers contain biologically
significant quantities of cations, e.g. Na" or K+. TRIS (Table 7.3) is often
toxic to biological systems: owing to its high lipid solubility it can penetrate
membranes, uncoupling electron transport reactions in whole cells and
isolated organelIes. In addition, it is markedly affected by temperature, with a
lO-fold increase in H+ concentration from 4 QC to 37"C. A number of
zwitterionic molecules (possessing both positive and negative groups) have been
introduced to overcome some of the disadvantages of the more traditional
buffers. These newer compounds are often referred to as 'Good buffers', to


acknowledge the early work of Dr N.B. Good and eo-workers: HEPES is one
of the most useful zwitterionic buffers, with a pKa of 7.5 at 25°C.
These zwitterionic substances are usually added to water as the free
acid: the solution must then be adjusted to the correct pH with a strong
alkali, usually NaOH or KOH. Alternatively, they may be used as their
sodium or potassium salts, adjusted to the correct pH with a strong acid,
e.g. HCI. Consequently, you may need to consider what effects such
changes in ion concentration may have in a solution where zwitterions are
used as buffers.
Fig. 7.4 shows a number of traditional and zwitterionic buffers and their
effective pH ranges. When selecting one of these buffers, aim for a pKa which
is in the direction of the expected pH change (Tables 7.2, 7.3). For example,
HEPES buffer would be a better choice of buffer than PIPES for use at

Table 7.3 pKa values at 25°C of some acids

pH 7.2 for experimental systems where a pH increase is anticipated, while
and bases (upper section) and some large PIPES would be a better choice for where acidification is expected.
organic zwitterions (lower section) commonly
used in buffer solutions. For polyprotic acids, Preparation of buffer solutions
where more than one proton my dissociate,
Having selected an appropriate buffer, you will need to make up your
the pKa values are given for each ionization
step. Only the trivial acronyms of the larger
solution to give the desired pH. You will need to consider two factors:
molecules are provided: their full names can 1. The ratio of acid and conjugate base required to give the correct pH.
be obtained from the catalogues of most 2. The amount of buffering required; buffer capacity depends upon the
chemical suppliers
absolute quantities of acid and base, as well as their relative proportions.
In most instances, buffer solutions are prepared to contain between
10mmol L -1 and 200 mmol L ~ 1 of the conjugate pair. While it is possible to
Acetic acid 4.8
Carbonic acid 6.1,10.2 calculate the quantities required from first principles using the Hendersori-
Citric acid 3.1, 4.8, 5.4 Hasselbalch equation, there are several sources which tabulate the amount of
Glycylglycine 3.1,8.2 substance required to give a particular volume of solution with a specific pH
Phthalic acid 2.9,5.5
Phosphoric acid 2.1,7.1,12.3 value for a wide range of traditional buffers (e.g. Perrin and Dempsey, 1974).
Succinic acid 4.2,5.6 For traditional buffers, it is customary to mix stock solutions of acidic and
TRIS* 8.3 basic components in the correct proportions to give the required pH (Table
Boric acid 9.2
7.4). For zwitterionic acids, the usual procedure is to add the compound to
MES 6.1
water, and then bring the solution to the required pH by adding a specific
PIPES 6.8 amount of strong alkali or acid (obtained from tables). Alternatively, the
MOPS 7.2 required pH can be obtained by dropwise addition of alkali or acid, using a
HEPES 7.5
meter to check the pH, until the correct value is reached. When preparing
TRICINE 8.1
TAPS 8.4 solutions of zwitterionic buffers, the acid may be relatively insoluble. Do not
CHES 9.3 wait for it to dissolve fully before adding alkali to change the pH - the
CAPS 10.4
addition of alkali will help bring the acid into solution (but make sure it has
all dissolved before the desired pH is reached).
*Note that this compound is hygroscopic and
should be stored in a desiccator.
Finally, when preparing a buffer solution based on tabulated information,
always confirm the pH with a pH meter before use.
Table 7.4 Preparation of sodium phosphate

buffer solutions for use at 25°C. Prepare
separate stock solutions of (a) disodium
hydrogen phosphate and (b) sodium
dihydrogen phosphate, both at 0.2 mol L~'.
Buffer solutions (at 0.1 mol L-1) are then
prepared at the required pH by mixing together
the volume of each stock solution shown in the
table, and then diluting to a final volume of
100mL using distilled or deionized water
4 6 10 11
6.0 6.2 43.8
pH
6.2 9.3 40.7
6.4 13.3 36.7 Fig. 7.4 Useful pH ranges of some commonly used buffers.
6.6 18.8 31.2
6.8 24.5 25.5
7.0 30.5 19.5
7.2 36.0 14.0
7.4 40.5 9.5
7.6 43.5 6.5
7.8 45.8 4.2
8.0 47.4 2.6

Resources
Resources for fundamental laboratory techniques
Books
Anon. (1989) Safe Practices in Chemical Laboratories, Royal Society of
Chemistry, London.
Anon. (1989) COSHH in Laboratories, Royal Society of Chemistry, London.
Bennett, S.W. and O'Neale, K. (1999) Progressive Development of Practical
Skills in Chemistry. A guide to early-undergraduate experimental work, Royal
Society of Chemistry, Cambridge.
Fumiss, BA, Hannaford, AJ., Smith, PW.G. and Tatchell, A.R. (1989)
Vogel's Textbook of Practical Organic Chemistry, 5th Edn, Longman,
Harlow, Essex.
Halpern, A.M. (1997) Experimental Physical Chemistry, Prentice Hall,
Harlow, Essex.
Harwood, L.M.,Moody, c.J. and Percy, J.M. (2000) Experimental Organic
Chemistry, 2nd Edn, Blackwell Science Ltd, Oxford.
Lehman, J.W. (1999) Operational Organic Chemistry. A problem-solving
approach to the laboratory course, 3rd Edn, Prentice Hall, Harlow, Essex.
Lenga R.E. (1988) Sigma-Aldricn Library of Chemical Safety Data, 2nd Edn,
Sigma-Aldrich Ltd, Gillingham.
Lister, T. (1996) Classic Chemistry Demonstrations, Royal Society of
Chemistry, Cambridge.
Mendham, J., Denney, R.C., Barnes, J.D. and Thomas, M.I.K. (2000)
Vogel's Textbook of Quantitative Chemical Analysis, 6th Edn, Prentice Hall,
Harlow, Essex.
Nelson, J.B. (1997) Laboratory Experiments for Chemistry, Prentice Hall,
Harlow, Essex.
Sharp, JT., Gosney, I. and Rowley, A.G. (1989) Practical Organic
Chemistry, Chapman and Hall, London.
Suib. S.L. and Tanaka, J. (1999) Experimental Methods in Inorganic
Chemistry, Prentice Hall, Harlow, Essex.
Urben, P.G. (1999) Bretherick's Handbook of Reactive Chemical Hazards, 6th
Edn, Butterworth-Heinemann, London.
Zubrick, J.W. (2001) The Organic Chem Lab Survival Manual. A student's
guide to techniques, 5th Edn, John Wiley and Sons Inc., Chichester.
Videos
Basic laboratory skills, LGC, Royal Society of Chemistry, Cambridge (1998).
Further Laboratory Skills, LGC, Royal Society of Chemistry, Cambridge
(1998).
Software
SoftCOSHH 2000, Royal Society of Chemistry, Cambridge.

------~8
Making and recording measurements
The term data (singular = datum, or data value or variate) refers to

measurements of a particular characteristic, or variable, classified as:
• Quantitative: where the individual values are described on a numerical scale
which may be either (i) continuous, taking any value on the measurement
scale, or (ii) discontinuous (or discrete), where only integer values are
possible. Many of the variables measured in chemistry are continuous and
quantitative, e.g. weight, temperature, time, amount of product formed in
an enzyme reaction.
• Ranked: where the data values can be listed in order of magnitude.
Where such data are given numbered ranks, they are sometimes called
'semi-quantitative data'. Note that such ranks cannot be treated as 'real'
numbers and they should not be added, averaged, etc.
• Qualitative: where individual values are assigned to a descriptive category,
e.g. the detection of the presence or absence of a chemical by a colour test
or precipitate.
Variables may be independent or dependent. Usually, the variable under the
control of the experimenter (e.g. time, reagent concentration, pH, etc.) is the
independent variable, while the variable being measured is the dependent
variable (p. 76). Sometimes, it is inappropriate to describe variables in this
way, and they are often referred to as interdependent. Another group of
values, often termed derived (or computed) data, are calculated from two or
more individual measurements, and these include ratios, percentages and
rates.
Measurement scales
Variables may be measured on different types of scale:
• Nominal - where classification is based on a descriptive characteristic
(e.g. colour). This is the only scale for qualitative data.
• Ordinal - this classifies by numerical rank, from smallest to greatest, but
with no assumption of equal spacing between ranks.
• Interval - for certain quantitative variables, where numbers on an equal
unit scale are related to an arbitrary zero, e.g. temperature in "C.
• Ratio - similar to the interval scale, except that the zero point represents
an absence of that character, i.e. it is an absolute zero.
You should aim to do quantitative measurements using a ratio scale
whenever possible, to allow you to use the broadest range of mathematical
operations and statistical procedures. For example, if you are measuring the
surface area of a sample of graphite you could give the area as 200 m2 but, if
you know the mass of the sample (10 g), you should quote the surface area as
20m2 g-l.
Accuracy and precision

Accuracy is the closeness of a measured or derived data value to its true
value, while precision is the closeness of repeated measurements to each other
(Fig. 8.1). A balance with a fault in it (i.e. a bias, see below) could give
precise (i.e. very repeatable) but inaccurate (i.e. untrue) results. Unless there
is bias in a measuring system, precision will lead to accuracy and it is
The investigative approach 65

precision that is generally the most important practical consideration, if there

is no reason to suspect bias. You can investigate the precision of any
precise measuring system by repeated measurements of the same sample: the nearer
but not
accurate
the replicate values are to each other, the more precise the measurement.
Absolute accuracy and precision are impossible to achieve, owing to the
limitations of measuring systems. It is particularly important to avoid
spurious accuracy in the presentation of results; include only those digits
which the accuracy of the measuring system implies. This type of error is
accurate common when changing units (e.g. inches to metres) and in derived data,
but not
precise
especially when calculators give results to a large number of decimal places.
Bias (systematic error)

Bias is a systematic or non-random inaccuracy and is one of the most
inaccurate troublesome difficulties in using numerical data. Biases may be associated
and with incorrectly calibrated instruments, e.g. a faulty electrode or syringe, or
imprecise
with experimental manipulations, e.g. decomposition of chemical compound
on storage. Bias in measurement can also be subjective, or personal, e.g. an
experimenter's preconceived ideas about an 'expected' result.
Bias can be minimized by using a carefully standardized procedure, with
precise fully calibrated instruments. Investigate bias in 'trial runs' by measuring a
and variable in several different ways, to see if the same result is obtained.
accurate
To avoid personal bias, 'blind' measurements should be made where the
identity of each sample is unknown to the operator, e.g. use a coding system.
Fig. 8.1 'Target' diagrams illustrating precision

and accuracy. Measurement (random) error
All measurements are subject to error, but the dangers of misinterpretation
are reduced by recognizing and understanding the likely sources of error and
by adopting appropriate protocols and calculation procedures.
A common source of random error is carelessness, e.g. reading a scale in
the wrong direction or parallax errors. This can be reduced greatly by careful
recording and may be detected by repeating the measurement. Other errors
arise from faulty or inaccurate equipment, but even a perfectly functioning
machine has distinct limits to the accuracy and precision of its measurements.
These limits are often quoted in manufacturers' specifications and are
applicable when an instrument is new; however, you should allow for some
deterioration with age.
One major influence virtually impossible to eliminate is the effect of the
investigation itself: even putting a thermometer in a liquid may change the
temperature of the liquid. The very act of measurement may give rise to a
confounding variable (p. 76) as discussed in Chapter 10. You should include
descriptions of the possible sources of error(s) and estimates of their
importance in any report and these should not be used as an excuse for poor
technique or inadequacies in your experimental design.
Collecting and recording primary data

When carrying out lab work or research projects, you will need to master
the important skills of recording and managing data. Individual observa-
tions (e.g. laboratory temperature) can be recorded in the text of your
notes, but tables are the most convenient way to collect large amounts of
information.
66 The investigative approach

~ A good set of lab notes should:
• outline the purpose of your experiment or observation;

• set down all the information required to describe your experimental
section;
• record all relevant information about your results or observations and
provide a visual representation of the data;
• note your immediate conclusions and suggestions for further experi-
ments.
When preparing a table for data collection, you should:

1. Use a concise title or a numbered code for cross-referencing.
2. Decide on the number of variables to be measured and their relationship
with each other and layout the table appropriately:
(a) The first column of your table should show values of the
independent (controlled) variable, with subsequent columns for the
individual (measured) values for each replicate or sample.
(b) If several variables are measured for the same organism or sample,
each should be given a row.
(c) In time-course studies, put the replicates as columns grouped
according to treatment, with the rows relating to different times.
3. Make sure the arrangement reflects the order in which the values will be
collected. Your table should be designed to make the recording process
as straightforward as possible, to minimize the possibility of mistakes.
For final presentation, a different arrangement may be best (Chapter 37).
4. Consider whether additional columns are required for subsequent cal-
culations. Create a separate column for each mathematical manipulation,
so the step-by-step calculations are clearly visible. Use a computer spread-
sheet (p. 307) if you are manipulating lots of data.
5. Use a pen to record data.
6. Take sufficient time to record quantitative data unambiguously - use
large, clear numbers, making sure that individual numerals cannot be
confused.
7. Record numerical data to an appropriate number of significant figures,
reflecting the accuracy and precision of your measurement (p. 65). Do
not round off data values, as this might affect the subsequent analysis.
8. Record the actual observations, not your interpretation, e.g. the colour
of a particular chemical test, rather than whether the test was positive or
negative. Take care not to lose any of the information content of the
data: for instance, if you only write down means and not individual
values, this will affect your ability to carry out subsequent statistical
analyses.
9. Prepare duplicated recording tables/checklists for repeated experiments.
10. Explain any unusual results in a footnote. Don't rely on memory.
Recording details of project work

The recommended system is one where you make a dual record.
Primary record
The primary record is made at the bench and you must concentrate on
the detail of materials, methods and results. Include information that
would not be used elsewhere, but which might prove useful in error
tracing: for example, if you note how a solution was made up (exact

volumes and weights used rather than concentration alone), this could
reveal whether a miscalculation had been the cause of a rogue result.
Note the origin, type and state of the chemicals used. In the experimental
section, the basic rule is to record enough information to allow a
reasonably competent scientist to repeat your work exactly. You must
tread a line between the extremes of pedantic, irrelevant detail and the
omission of information essential for proper interpretation - better
perhaps to err on the side of extra detail to begin with. An experienced
worker can tell you which subtle shifts in technique are important (e.g.
batch numbers for an important chemical, or when a new stock solution
is prepared). Many important scientific advances have been made because
of careful observation and record taking and because coincident data
were recorded that did not seem of immediate value. Make rough
diagrams to show the arrangement of replicates, equipment, etc. If forced
to use loose paper to record data, make sure each sheet is dated and
taped to your lab book, collected in a ring binder, or attached with a
treasury tag. The same applies to traces, printouts and graphs.
The basic order of the primary record should mirror that of a research
report (see p. 332), including: the title and date, brief introduction, a
comprehensive experimental section, the data and short conclusions.
Secondary record
You should make a secondary record concurrently or later in a bound
book and it ought to be neater, in both organization and presentation.
This book will be used when discussing results with your supervisor, and
when writing up a report or thesis, and may be part of your course
assessment. Writing a second, neater version forces you to consider again
details that might have been overlooked in the primary record and
provides a duplicate in case of loss or damage. While these notes should
retain the essential features of the primary record, they should be more
concise and the emphasis should move towards analysis of the experi-
ment. Don't repeat the experimental section for a series of similar
experiments; use devices such as 'method as for Expt B4'. A photocopy
may be sufficient if the method is derived from a text or article (check
with your supervisor). Outline the aims more carefully at the start and
link the experiment to others in a series (e.g. 'Following the results of
Expt D24, I decided to test whether .. .'). You should present data in an
easily digested form, e.g. as tables of means or as summary graphs. Use
appropriate statistical tests (p. 271) to support your analysis of the
results. Always analyse and think about data immediately after collecting
them as this may influence your subsequent activities. Write down any
conclusions: sometimes those which seem obvious at the time of doing the
work are forgotten when the time comes to write up a report or thesis.
Likewise, ideas for further studies may prove valuable later. Even if your
experiment appears to be a failure, suggestions as to the likely causes
might prove useful.
Using communal records

If working with a research team, you may need to use their communal
databases. These avoid duplication of effort and ensure uniformity in
techniques. They may also form part of the legal safety requirements for lab
work. They might include:

• a shared notebook of common techniques (e.g. solutions or calibration

technique);
• a set of simplified step-by-step instructions for use of equipment;
• an alphabetical list of suppliers of equipment and consumables;
• a list of chemicals required by the group and where they are stored;
• the risk assessment sheets for dangerous procedures (p. 7);
• a record of the use and disposal of chemicals and solvents.

-0) SI units and their use
When describing a measurement, you normally state both a number and a

unit (e.g. 'the length is 1.85 metres'). The number expresses the ratio of
the measured quantity to a fixed standard, while the unit identifies that
standard measure or dimension. Clearly, a single unified system of units is
essential for efficient communication of such data within the scientific
community. The Systeme International d'Unites (SI) is the internationally
ratified form of the metre-kilogram-second system of measurement and
Table 9. 1 The base and supplementary
represents the accepted scientific convention for measurements of physical
SI units
quantities.
Another important reason for adopting consistent units is to simplify
complex calculations where you may be dealing with several measured
Base units
quantities (see p. 260). Although the rules of the SI are complex and the scale
Length metre m
Mass kilogram kg of the base units is sometimes inconvenient, to gain the full benefits of the
Amount of system you should observe its conventions strictly.
substance mole mol
Time s
The description of measurements in SI involves:
second
Electric current ampere A
Temperature kelvin K
• seven base units and two supplementary units, each having a specified
Luminous intensity candela cd abbreviation or symbol (Table 9.1);
Supplementary units • derived units, obtained from combinations of base and supplementary
Plane angle radian rad units, which may also be given special symbols (Table 9.2);
Solid angle steradian sr • a set of prefixes to denote multiplication factors of 103, used for
convenience to express multiples or fractions of units (Table 9.3).
Table 9.2 Some important derived SI units
Energy joule J m2 kgs-2 Nm

Force newton N m kgs-2 J rn"
Pressure pascal Pa kg m-1 S-2 N m-2
Power watt W m2 kg S-3 J S-l
Electric charge coulomb C As J V-1
Electric potential
Table 9.3 Prefixes used in the SI difference volt V m2 kg A-1 S-3 J C-1
Electric resistance ohm n m2 kg A-2 S-3 V A-1
Electric conductance siemens S S3 A2 kg-1 m-2 A V-1 or n-1
Electric capacitance farad F S4 A2 kg-1 m-2 C V-1
10-3 milli m 103 kilo k
10-6 Luminous flux lumen Im cd sr
micro J1 106 mega M
10-9 Illumination lux Ix cd sr m-2 Im m-2
nano n 109 giga G
Frequency hertz Hz S-l
10-12 pi eo p 1012 tera T
10-15 femto f 1015 peta P Radioactivity becquerel Bq s"
10-18 atto a 1018 exa E Enzyme activity katal kat mol substrate S-l
Recommendations for describing measurements in SI units

Basic format
• Express each measurement as a number separated from its units by a
space. If a prefix is required, no space is left between the prefix and the
unit it refers to. Symbols for units are only written in their singular form
and do not require full stops to show that they are abbreviated or that
they are being multiplied together.

SI units and their use
• Give symbols and prefixes appropriate upper or lower case initial letters
as this may define their meaning. Upper case symbols are named after
persons but when written out in full they are not given initial capital
letters.
• Show the decimal sign as a full point on the line. Some metric
countries continue to use the comma for this purpose and you may
come across this in the literature: commas should not therefore be used
to separate groups of thousands. In numbers that contain many
significant figures, you should separate multiples of 103 by spaces
rather than commas.
Compound expressions for derived units

• Take care to separate symbols in compound expressions by a space to
avoid the potential for confusion with prefixes. Note, for example, that
200 m s (metre seconds) is different from 200 ms (milliseconds).
• Express compound units using negative powers rather than a solidus U):
for example, write mol m-3 rather than mol/m". The solidus is reserved
for separating a descriptive label from its units (see p. 251).
• Where there is a choice, select relevant (natural) combinations of
derived and base units, e.g. you might choose units of Pa m " to
describe a hydrostatic pressure gradient rather than kg m ? s", even
though these units are equivalent and the measurements are numerically
the same.
Use of prefixes
• Use prefixes to denote multiples of 103 (Table 9.3) so that numbers are
kept between 0.1 and 1000.
• Treat a combination of a prefix and a symbol as a single symbol. Thus,
when a modified unit is raised to a power, this refers to the whole unit
including the prefix.
• Avoid the prefixes deci (d) for 10-1 and centi (c) for 10-2 as they are not
strictly SI.
• Express very large or small numbers as a number between 1 and 10
multiplied by a power of 10 if they are outside the range of prefixes
shown in Table 9.3.
• Do not use prefixes in the middle of derived units: they should be
attached only to a unit in the numerator (the exception is in the unit for
mass, kg).
~ For the foreseeable future. you will need to make con-

versions from other units to SI units. as much of the literature quotes
data using imperial. c.g.s. or other systems. You will need to recognize
these units and find the conversion factors required. Examples relevant
to chemistry are given in Box 9.1. Table 9.4 provides values of some im-
portant physical constants in SI units.
Some implications of SI in chemistry

Volume
The SI unit of volume is the cubic metre, m", which is rather large for
practical purposes. The litre (L) and the millilitre (mL) are technically
obsolete, but are widely used and glassware is still calibrated using them.

Table 9.4 Some physical constants in SI terms
Avogadro's constant 6.022174 X 1023 rnol "

Boltzmann's constant 1.380626J K-l
Charge of electron 1.602192 x 10-19 C
Gas constant 8.31443J K-l rnol'
Faraday's constant 9.648675 x 104Cmol-1
Molar volume of ideal gas at STP 0.022414 m3 mot'
Speed of light in vacuo 2.997924 x 108 m S-l
Planck constant 6.626205 x 10-34 J s
Acceleration due to gravity 9.807 m S-l
Atomic mass unit 16605402 x 10-27 kg
Rydberg constant 1.097 x 107 rn"
Permitivity of vacuum 8.854 x 10-12Fm-1

Mass
The SI unit for mass is the kilogram (kg) rather than the gram (g): this is
unusual because the base unit has a prefix applied.
Amount of substance
You should use the mole (mol, i.e. Avogadro's constant, see Table 9.4) to
express very large numbers. The mole gives the number of atoms in the
atomic mass, a convenient constant.
Concentration
The SI unit of concentration, mol m>', is not convenient for general
laboratory work. It is equivalent to the non-SI term 'millimolar' (mM) while
'molar' (M) becomes kmol m ". If the solvent is not specified, then it is
assumed to be water (see Chapter 6).
Time
In general, use the second (s) when reporting physical quantities having a
time element. Hours (h), days (d) and years should be used if seconds are
clearly absurd (e.g. samples were taken over a 5-year period). Note,
however, that you may have to convert these units to seconds when doing
calculations.
Temperature
The SI unit is the kelvin, K. The degree Celsius scale has units of the
same magnitude, QC, but starts at 273.15 K, the melting point of ice at
STP. Temperature is similar to time in that the Celsius scale is in
widespread use, but note that conversions to K may be required for
calculations. Note also that you must not use the degree sign (0) with K
and that this symbol must be in upper case to avoid confusion with k for
kilo; however, you should retain the degree sign with QC to avoid
confusion with the coulomb, C.

Interconversion of SI units
You will find that the use of SI units simplifies mathematical manipulations
and ensures that you obtain the correct units for the parameter being
calculated. Remember that you must convert all units into the appropriate SI
units, e.g. masses must be expressed as kg, volumes as m3 and concentrations
as kg m-3 or mol m-3, etc., and that you may need to use alternatives in
derived units (Table 9.2). The application of these principles is shown in Box
9.2.

Scientific method and design of
experiments
Science is a systematized body of knowledge derived from observation and

experiment. Scientists carry out experiments, make observations and attempt
to explain the results; these tentative explanations are called hypotheses and
their validity is tested by systematically forming and rejecting alternative
explanations.
Some branches of chemistry are designed to provide fundamental
information, rather than to test a particular hypothesis: for example, the
purification and characterization of a newly discovered naturally occurring
molecule. In contrast, an experiment is a contrived situation designed to test
one or more hypothesis under conditions controlled by the investigator. Any
hypothesis that cannot be rejected from the results of an experiment is
provisionally accepted. This 'sieve' effect leaves us with a set of current
explanations for our observations. These explanations are not permanent and
may be rejected on the basis of a future investigation. A hypothesis that has
withstood many such tests and has been shown to allow predictions to be
made is known as a theory, and a theory may generate such confidence
through its predictive abilities to be known as a law (Fig. 10.1).
Observations are a prelude to experimentation, but they are preconditioned
by a framework of peripheral knowledge. While there is an element of luck in
being at the right place and time to make important observations, as Pasteur
stated, 'chance favours only the prepared mind'. A fault in scientific method is
that the design of the experiment and choice of method may influence the
outcome - the decisions involved may not be as objective as some scientists
assume. Another flaw is that radical alternative hypotheses may be overlooked
in favour of a modification to the original hypothesis, and yet just such leaps
in thinking have frequently been required before great scientific advances.
No hypothesis can ever be rejected with certainty. Statistics allow us to
quantify as vanishingly small the probability of an erroneous conclusion, but
we are nevertheless left in the position of never being 100% certain that we
have rejected all relevant alternative hypotheses, nor 100% certain that our
decision to reject some alternative hypotheses was correct! However, despite
these problems, experimental science has yielded and continues to yield many
important findings.
~ The fallibility of scientific 'facts' is essential to grasp.

No explanation can ever be 100% certain as it is always possible for a
new alternative hypothesis to be generated. Our understanding of
chemistry changes all the time as new observations and methods force
old hypotheses to be retested. .
Fig. 70.7 How scientific investigations proceed.
Quantitative hypotheses involve a mathematical description of the system.
They can be formulated concisely by mathematical models. Models are often
useful because they force deeper thought about mechanisms and encourage
simplification of the system. A mathematical model:
• is inherently testable through experiment;

• identifies areas where information is lacking or uncertain;
• encapsulates many observations;
• allows you to predict the behaviour of the system.

Scientific method and design of experiments
Remember, however, that assumptions and simplifications required to

create a model may result in it being unrealistic. Further, the results obtained
from any model are only as good as the information put into it.
Experimentation and variables

In many experiments, the aim is to provide evidence for causality. If x causes
y, we expect, repeatably, to find that a change in x results in a change in y.
Hence, the ideal experiment of this kind involves measurement of y, the
dependent (measured) variable, at one or more values of x, the independent
variable, and subsequent demonstration of some relationship between them.
Experiments therefore involve comparisons of the results of treatments -
changes in the independent variable as applied to an experimental subject.
The change is engineered by the experimenter under controlled conditions.
Experimental subjects given the same treatment are known as replicates.
Interpretation of experiments is seldom clear-cut because uncontrolled
variables always change when treatments are given.
Confounding variables
These increase or decrease systematically as the independent variable in-
creases or decreases. Their effects are known as systematic variation. This
form of variation can be disentangled from that caused directly by treatments
by incorporating appropriate controls in the experiment. A control is really
just another treatment where a potentially confounding variable is adjusted
so that its effects, if any, can be taken into account. The results from a
control may therefore allow an alternative hypothesis to be rejected. There
are often many potential controls for any experiment.
The consequence of systematic variation is that you can never be certain
that the treatment, and the treatment alone, has caused an observed result.
By careful design, you can, however, 'minimize the uncertainty' involved in
your conclusion. Methods available include:
• Ensuring, through experimental design, that the independent variable is
the only major factor that changes in any treatment.
• Incorporating appropriate controls to show that potential confounding
variables have little or no effect.
• Selecting experimental subjects randomly to cancel out systematic
variation arising from biased selection.
• Matching or pairing individuals among treatments so that differences in
response due to their initial status are eliminated.
• Arranging subjects and treatments randomly so that responses to
systematic differences in conditions do not influence the results.
• Ensuring that experimental conditions are uniform so that responses to
systematic differences in conditions are minimized.
Nuisance variables
These are uncontrolled variables which cause differences in the value of y
independently of the value of x, resulting in random variation. Nuisance
variables are not common in chemistry except where molecules from natural
sources are used. To reduce and assess the consequences of nuisance
variables:
• incorporate replicates to allow random variation to be quantified;
• choose experimental subjects that are as similar as possible;
• control random fluctuations in environmental conditions.

Constraints on experimental design

Box 10.1 outlines the important stages in designing an experiment. At an
early stage, you should find out how your resources may constrain the
design. For example, limits may be set by availability of subjects, cost of
treatment, availability of a chemical or bench space. Logistics may be a
factor (e.g. time taken to record or analyse data), or your equipment and
facilities may affect design because you cannot regulate conditions as well as
you might desire. You should also consider what statistical tests you intend
to make (p. 271), as this is an important part of experimental design.
Using replicates
Replicate results show how variable the response is within treatments. They
allow you to compare the differences among treatments in the context of the
variability within treatments - you can do this via statistical tests such as
analysis of variance (Chapter 41). Larger sample sizes tend to increase the
precision of estimates of statistical parameters and increase the chances of
showing a significant difference between treatments if one exists. For
statistical reasons (weighting, ease of calculation, fitting data to certain tests),
it is best to keep the number of replicates even.

3x3 4x4 If the total number of replicates available for an experiment is limited by
resources, you may need to compromise between the number of treatments
and the number of replicates per treatment. Statistics can help here, as it is
possible to work out the minimum number of replicates you would need to
show a certain difference between pairs of means (say 10%) at a specified
level of significance (say P = 0.05). For this, you need to obtain a prior
estimate of variability within treatments (see Miller and Miller, 2000).
Fig. 10.2 Examples of Latin square Randomization of treatments

arrangements for three and four treatments.
Letters indicate treatments; the number of For relatively simple experiments, you can adopt a completely randomized
possible arrangements increases greatly as the design; here, the position and treatment assigned to any subject is defined
number of treatments increases. randomly. You can draw lots, use a random number generator on a
calculator, or use the random number tables which can be found in most
books of statistical tables.
A completely randomized layout has the advantage of simplicity but
cannot show how confounding variables alter in space or time. This
information can be obtained if you use a blocked design in which the degree
of randomization is restricted. Here, the experimental space or time is divided
into blocks, each of which accommodates the complete set of treatments.
When analysed appropriately, the results for the blocks can be compared to
test for differences in the confounding variables and these effects can be
natural water separated out from the effects of the treatments. The size and shape (or
timing) of the block you choose is important: besides being able to
3 4
accommodate the number of replicates desired, the suspected confounding
variable should berelatively uniform within the block.
A Latin square is a method of placing treatments so that they appear in a
balanced fashion within the experimental area. Treatments appear once in
each column and row (see Fig. 10.2), so the effects of confounding variables
can be 'cancelled out' in two directions at right angles to each other. This is
effective if there is a smooth gradient in some confounding variable over the
experimental area. It is less useful if the variable has a patchy distribution,
where a randomized design might be better.
Fig. 10.3 The experimenter wishes to Latin square designs are useful in serial experiments where different
investigate the effect of pH A-E on the treatments are given to the same subjects in a sequence (e.g. Fig. 10.3). A
extraction of phenols from five natural waters.
disadvantage of Latin squares is the fact that the number of columns and
Each water sample is extracted at a different pH
and the influence of pH on the recovery of rows is equal to the number of replicates, so increases in the number of
phenols noted. replicates can only be made by the use of further Latin squares.
Multifactorial experiments
The simplest experiments are those in which one treatment (factor) is applied
at a time to the samples. This approach is likely to give clear-cut answers, but
it could be criticized for lacking realism. In particular, it cannot take account
of interactions among two or more conditions that are likely to occur in real
life. A multifactorial experiment (Fig. lOA) is an attempt to do this; the
interactions among treatments can be analysed by specialized forms of
analysis of variance.
Multifactorial experiments are economical on resources because of 'hidden
replication'. This arises when two or more treatments are given to a subject
because the result acts statistically as a replicate for each treatment. Choice of
relevant treatments to combine is important in multifactorial experiments; for
instance, an interaction may be present at certain concentrations of a
chemical but not at others (perhaps because the response is saturated). It is

factor B
+
factor
A
Fig. 10.4 Design of a simple multifactorial experiment. Factors A and B have

effects a and b when applied alone. When both are applied together, the effect is
denoted by a + b + c.
• If c = 0, there is no interaction
(e.g. 2 + 2 + c = 4).
• If c is positive, there is a positive interaction (synergism) between A and B
(e.g. 2 + 2 + c = 5).
• If c is negative, there is a negative interaction (antagonism) between A and B
(e.g. 2 + 2 + c = 3).
also important that the measurement scale for the response is consistent,
otherwise spurious interactions may occur. Beware when planning a
multifactorial experiment that the numbers of replicates do not get out of
hand: you may have to restrict the treatments to 'plus' or 'minus' the factor
of interest (as in Fig. 10.4).
Repetition of experiments
Even if you have taken great care to ensure that your experiment is well
designed and statistically analysed, you are limited in the conclusions that can
be made. Firstly, what you can say is valid for a particular place and time,
with a particular investigator, experimental subject and method of applying
treatments. Secondly, if your results were significant at the 5% level of
probability, there is still an approximately 1 in 20 chance that the results did
arise by chance. To guard against these possibilities, it is important that
experiments are repeated. Ideally, this would be done by an independent
scientist with independent materials. However, it makes sense to repeat work
yourself so that you can have full confidence in your conclusions. Many
scientists recommend that all experiments are carried out three times in total.
This may not be possible in undergraduate practical classes or project work!

------~8 Project work
Research projects are an important component of the final-year syllabus for

most degree programmes in chemistry, while shorter projects may also be
carried out during courses in earlier years. Project work presents difficulties
at many stages but can be extremely rewarding. The assessment of your
project is likely to contribute significantly to your degree grade, so all aspects
of this work should be approached in a thorough manner.
Deciding on a topic to study

Assuming you have a choice, this important decision should be researched
carefully. Make appointments to visit possible supervisors and ask them for
advice on topics that you find interesting. Use library texts and research
papers to obtain further background information. Perhaps the most
important criterion is whether the topic will sustain your interest over the
whole period of the project. Other things to look for include:
• Opportunities to learn new skills. Ideally, you should attempt to gain
experience and skills that you might be able to 'sell' to a potential
employer.
• Ease of obtaining valid results. An ideal project provides a means to
obtain 'guaranteed' data for your report, but also the chance to extend
knowledge by doing genuinely novel research.
• Assistance. What help will be available to you during the project? A busy
lab with many research students might provide a supportive environment
should your potential supervisor be too busy to meet you often; on the
other hand, a smaller lab may provide the opportunity for more personal
interaction with your supervisor.
• Impact. It is not outside the bounds of possibility for undergraduate
work to contribute to research papers. Your prospective supervisor can
alert you to such opportunities.
• Success. You are doing a research project and it may not always provide
a positive result. Negative results are just as useful.
Planning your work

As with any lengthy exercise, planning is required to make the best use of the
time allocated. This is true on a daily basis as well as over the entire period
of the project. It is especially important not to underestimate the time it will
take to write and produce your thesis (see below). If you wish to benefit from
feedback given by your supervisor, you should aim to have drafts in his/her
hands in good time. Since a large proportion of marks will be allocated to
the report, you should not rush its production.
If your department requires you to write an interim report, look on this
as an opportunity to clarify your thoughts and get some of the time-
consuming preparative work out of the way. If not, you should set your
own deadlines for producing drafts of the introduction, materials and
methods section, etc.
Project work can be very time consuming at times. Try not to neglect other
aspects of your course - make sure your lecture notes are up to date and
collect relevant supporting information as you go along.

Project work
Getting started
Fig. 11.1 is a flowchart illustrating how a project might proceed; at the start,
don't spend too long reading the literature and working out a lengthy
programme of research. Get stuck in and do an experiment. There's no
substitute for 'getting your hands dirty' for stimulating new ideas:
• even a 'failed' experiment will provide some useful information which

may allow you to create a new or modified hypothesis;
• pilot experiments may point out deficiencies in experimental technique
that will need to be rectified;
• the experience will help you create a realistic plan of work.
Designing experiments or sampling procedures

Design of experiments is covered in Chapter 10. Avoid being too ambitious
at the start of your work! It is generally best to work with a simple
hypothesis and design your experiments or sampling around this. A small
pilot experiment or test sample will highlight potential stumbling blocks
including resource limitations, whether in materials or time or both.
Working in a laboratory environment

During your time as a project student, you are effectively a guest in your
supervisor's laboratory.
Fig. 11.1 Flowchart showing a recommended • Be considerate - keep your 'area' tidy and offer to do your share of lab
sequence of events in carrying out an duties such as calibrating the pH meter, replenishing stock solutions,
undergraduate research project. distilled water, cleaning used glassware, etc.
• Use instruments carefully - they could be worth more than you'd think.
Careless use may invalidate calibration settings and ruin other people's
work as well as your own.
• Do your homework on techniques you intend to use - there's less chance
of making costly mistakes if you have a good background understanding
of the methods you will be using.
• Always seek advice if you are unsure of what you are doing.
~ It is essential that you follow all the safety rules ap-

plying to the laboratory or field site. Make sure you are acquainted with
all relevant procedures - normally there will be prominent warnings
about these. If in doubt, ask!
Keeping notes and analysing your results

Tidy record keeping is often associated with good research, and you should
follow the advice and hints given in Chapter 8. Try to keep copies of all files
relating to your project. As you obtain results, you should always calculate,
analyse and graph data as soon as you can (see Fig. 11.1). This can reveal
aspects that may not be obvious in numerical or readout form. Don't be
worried by negative results - these can sometimes be as useful as positive
results if they allow you to eliminate hypotheses - and don't be dispirited if
things do not work first time. Thomas Edison's maxim 'Genius is one per
cent inspiration and ninety-nine per cent perspiration' certainly applies to
research work!

Project work
Writing the report

The structure of scientific reports is dealt with in Chapter 52. The following
advice concerns methods of accumulating relevant information.
Introduction This is a big piece of writing that can be very time-

consuming. Therefore, the more work you can do on it early on, the better.
You should allocate some time at the start for library work (without
neglecting benchwork), so that you can build up a database of references (p.
319). While photocopying can be expensive, you will find it valuable to have
copies of key reviews and references handy when writing away from the
library. Discuss proposals for content and structure with your supervisor to
make sure your effort is relevant. Leave space at the end for a section on
aims and objectives. This is important to orientate readers (including
assessors), but you may prefer to finalize the content after the results have
been analysed!
Experimental You should note as many details as possible when doing the
experiment or making observations. Don't rely on your memory or hope that
the information will still be available when you come to write up. Even if it
is, chasing these details might waste valuable time.
Results Show your supervisor graphed and tabulated versions of your data
promptly. These can easily be produced using a spreadsheet (p. 307), but you
should seek your supervisor's advice on whether the design and print quality
is appropriate to be included in your thesis. You may wish to access a
specialist graphics program to produce publishable-quality graphs and charts:
allow some time for learning its idiosyncrasies! If you are producing a poster
for assessment (Chapter 54), be sure to mock up the design well in advance.
Similarly, think ahead about your needs for any seminar or poster you will
present.
Discussion Because this comes at the end of your thesis, and some parts can
only be written after you have all the results in place, the temptation is to leave
the discussion to last. This means that it can be rushed - not a good idea
because of the weight attached by assessors to your analysis of data and
thoughts about future experiments. It will help greatly if you keep notes of
aims, conclusions and ideas for future work as you go along (Fig. 11.1).
Another useful tip is to make notes of comparable data and conclusions from
the literature as you read papers and reviews.
Acknowledgements Make a special place in your notebook for noting all

those who have helped you carry out the work, for use when writing this
section of the report.
References Because of the complex formats involved (p. 319), these can be
tricky to type. To save time, process them in batches as you go along.
~ Make sure you are absolutely certain about the dead-

line for submitting your report and try to submit a few days before it. If
you leave things until the last moment, you may find access to printers,
photocopiers and binding machines is difficult.

Resources
Resources for the investigative approach
Books
Adams, M.J. (1995) Chemometrics in Analytical Chemistry, Royal Society of
Crawford, K. and Heaton, A. (1999) Problem Solving in Analytical
Chemistry, Royal Society of Chemistry, Cambridge.
Massart, D.L., Vandeginste, B.G.M., Buydens, L.M.C, de Jong, S., Lewi,
PJ. and Smeyers-Verbeke, J. (1997) Handbook of Chemometrics and
Qualimetrics: Part A, Elsevier Science RV., Amsterdam.
Meier, P.C (2000) Statistical Methods in Analytical Chemistry, 2nd Edn,
John Wiley and Sons Ltd, Chichester.
Miller, J.N. and Miller, J.C (2000) Statistics and Chemometrics for Analytical
Chemistry, 4th Edn, Prentice Hall, Harlow, Essex.

------~0
Melting points
Melting points are measured (determined) for four reasons:
1. The melting range and upper limit are an indication of the purity of the
sample.
2. Comparison of the melting point with the literature may indicate the
identity of the compound or confirm that it is not the compound required.
3. If the compound is new, other scientists will need the information.
4. The compound can be identified with reasonable certainty by taking a
mixed melting point (p. 90).
Criterion of purity
Pure solid covalent organic compounds and many inorganic complexes
incorporating organic ligands have definite melting points. The pure solid will
melt reproducibly over a narrow temperature range, usually less than 1 QC,and
this melting range is known as the melting point. If the compound is not pure,
the melting range will increase significantly and the upper end of the melting
range will be lowered. Thus the melting point (m.pt.) of a compound is a
measure of its purity (p. 90). Other methods are used routinely to estimate
purity of solid compounds, such as NMR (p. 190), and the presence of a single
'spot' or a single peak on a chromatogram from thin-layer (p. 216), gas-liquid
(p. 211) or high-performance liquid (p. 218) chromatography, but melting point
remains the standard measure of purity.
~ The term melting point really means the melting

range of a chemical and in your laboratory report you should always
quote the measured melting range under the heading 'melting point'.
Melting point apparatus

The equipment for measuring the melting point of a solid varies in
complexity from a simple oil bath heated with a micro burner to a microscope
with a heated stage as shown in Fig. 12.1. The essential components of a
melting point apparatus are:
• A sample holder: usually a glass capillary tube sealed at one end in the
case of the oil bath and heated block systems, or a pair of microscope
slides on an electrically heated plate in the Kofler block.
• A temperature recording device, placed as near to the sample as possible.
Usually this is a thermometer but it can be a thermocouple probe with
digital readout.
• A heat source with fine control to allow a gradual increase in tempera-
ture. These sources vary with the sophistication of the equipment.
In the undergraduate laboratory you are likely to use only the simpler
systems such as the oil bath or heated block apparatus.
Capillary tubes
Capillary tubes for melting point measurement are available commercially
and are supplied open at both ends or closed at one end or closed at both
Laboratory techniques 87
Melting points
(h)
(a)
(c) (d)
Fig. 12.1 Melting point apparatus: (a) oil bath; (b) heated block - thermometer
readout; (c) heated block - digital readout; (d) Kofler hot-stage microscope.
ends. To seal one end of an open capillary tube, just touch the end of the
capillary tube onto the outer 'layer' of the hot flame of a microburner (see
Fig. 12.2). The end of the tube will collapse in and seal the tube. Make sure
that the tube is sealed, i.e. there is not a fine line in the sealed end, and that
there is no large globule of glass on the end of the tube, otherwise it may not
fit into the hole in the heating block of the melting point apparatus.
Similarly, if you push the tube too far into the flame, the tube will bend and
therefore not fit into the heating block.
To put the compound into the capillary tube, place a little of the dry
compound in a small heap on a watch-glass and press the open end of the tube
into the heap, trapping a plug of chemical in the opening. The chemical can be
88 Laboratory techniques
Melting points
moved to the sealed end by turning the tube over and tapping it on the bench,
==="':>
? or by vibrating it by rubbing it against the thread of the screw on a clamp, or
dropping it down a long glass tube onto the laboratory bench. Remember: you
==,,0
WRONG!
only need 2-3 mm of sample in the bottom of the capillary tube.
Tube has been
overheated
tube tilted to stages for Oil bath apparatus
prevent water correct sealing
condensation inside
The component parts of a typical oil bath melting point apparatus are shown
in Fig. 12.3.
Fig. 12.2 Sealing a melting point capillary
tube. • Check that the mineral oil is clean and contains no water (p. 34) and that
the bulb is only two-thirds full, to allow for expansion. Clamp the oil
bath to a support stand (p. 26).
• Check that your thermometer is of the appropriate range, that the mercury
thread is intact and the glass, in particular the bulb, is not cracked.
• Attach the capillary to the thermometer with a rubber ring, making sure
that the compound is next to the thermometer bulb and remember to
hold the thermometer near the bulb while attaching the capillary tube
(p. 13).
• Press the thermometer into the 'notched' cork, making sure that you can
see the thermometer scale, and trial fit the thermometer, sample and cork
into the oil bath making sure that the thermometer bulb and sample are
in the centre of the oil bath and that the rubber band is not in the oil,
and will not be covered by oil, when the oil expands on heating.
• If adjustment is needed, carefully slide the cork up or down the thermo-
meter (p. 13).
• You can attach a melting point capillary to both sides of the thermometer
bulb to carry out two separate melting point determinations simul-
taneously.
Electric heated block apparatus

split for ~
The heating block usually has three holes, to permit three simultaneous
viewing rubber
m.pt.~
measurements of melting points. Always ensure that:
thermometer band
tube
• The heating block is at room temperature at the outset.
liquid 1sample
paraffin _ size • The light in the heating block works.
bath t 2-3 mm • The thermometer is undamaged (as above) and fits snugly into its hole in
the heating block.
~ microburner
• All heating controls are set at zero.
• The capillary tubes slide easily into their holes in the heating block.
Fig. 12.3 Components of an oil bath melting
point apparatus.
The usual problems encountered with this equipment are broken capillary
tubes or broken thermometer bulbs in the heating block. Consult your
instructor in these cases.
Melting point determination

The general procedure is to heat the oil bath with the micro burner using
the technique described on p. 32, or to use the heating control to give a
temperature rise of about 10 QC per minute. When the temperature is
about 20 QC below the melting point, the rate of heating must be reduced
to about 2 QC per minute and continued at this rate until the compound
has melted. The melting point is the temperature range from where the
first drop of liquid appears to where the last crystal dissolves into the
liquid.
Melting points
The following points of technique should be considered:

• The most common error is heating the sample too quickly. There is often
a lag between slowing the heating rate and the reduction in temperature
increase, resulting in a reading of the melting point which is too high.
• If you do not know what the compound is, you will not know its
expected melting point. Therefore, you should determine an approximate
value and then repeat the procedure to give an accurate measurement, by
reducing the heating rate, particularly for the final stage near the melting
point. The boost heater on the electrical apparatus can be used for
finding the approximate melting point but not for accurate determination.
• If you are carrying out several melting point measurements, it is common
sense to measure the melting point of the lowest melting compound first -
less time for the apparatus to cool to room temperature.
• If you miss the melting point, do not allow the sample to solidify and
then retake the melting point with the same sample: some decomposition
may have occurred on melting.
Thermometer calibration
Your thermometer may be a major source of error in melting point
measurement. Occasionally thermometers for routine laboratory use may not
be accurate and may read up to 10 DC high or low. To avoid this problem
you should always calibrate your thermometer to determine any error and be
able to correct for it.
To calibrate your thermometer you must measure the melting points of a
series of very pure compounds, available commercially, having a range of
melting points similar to the range over which you will use the thermometer;
a general-purpose series is shown in Table 12.1. Having measured the melting
points of each of the pure compounds, take the mid-point of each value for
the thermometer reading of melting point for each compound and the mid-
point of the literature melting point of each compound and plot them
Table 12.1 A typical series of standards for graphically using literature temperature as the y-axis and the thermometer
thermometer calibration reading as the x-axis, The straight-line plot will obey the equation
y = mx + c, where y = real temperature (literature m.pt.), x = thermometer
reading, m = slope and c = intercept. If you use a computer and suitable
Naphthalene 79-80°C program, the values of m and c will be calculated and you then solve the
Benzoic acid 121°C equation for using the thermometer reading (x-axis values) to find the real
4-Nitroaniline 147°C
4-Toluic acid 180-182"C temperature of melting (y-axis values) to find the true melting range.
Anthracene 216-218°C Of course, if you break or lose your thermometer, you must calibrate
another.
Mixed melting point determination

You can confirm the identity of a compound by determining a mixed melting
point. If you prepare a mixture of your unknown chemical and the one you
suspect it may be and measure the melting point of the mixture then there are
two possible results:
1. The melting point of the mixture is the same as the pure compound,
which means that the unknown compound and the known compound
are the same.
Melting points
2. The melting point of the mixture is lower than either of the two pure
components and the melting range is large. This is because the two
compounds are different with the result that one is an impurity in the
other.
For example, both benzoic acid and mandelic acid are white crystalline
solids which melt at 121°C. However a 1: 1 mixture of the two compounds
begins to melt at about 80°C.
The usefulness of mixed melting points is limited in that you must have
some idea of the chemical nature of your unknown compound and a sample
of the suspected compound must be available.
-------0 Recrystallization
The products from many synthetic preparations are seldom pure and the
technique of recrystallization, which involves dissolving the impure material
in a hot solvent and then cooling the solution to produce crystals, is routinely
used to purify covalent organic and inorganic solids.
In general there are three types of impurities, which are removed by the
recrystallization process:
1. Insoluble material: anti-bumping granules, pieces of filter paper, traces of

drying agents, grit, hair and other materials which may have been
present in the starting chemicals.
2. Small quantities of unreacted starting chemicals and/or by-products from
side reactions or other isomers.
3. Very small amounts of coloured by-products resulting from oxidation or
polymerization of the chemicals used.
Recrystallization is designed to remove all these types of impurity and
provide a pure product suitable for melting point measurement. Purification
by recrystallization is based on the theory of saturated solutions (p. 50) and a
suitable recrystallization solvent is one in which the chemical to be purified is
insoluble in the cold solvent and soluble in the hot solvent.
When the crude reaction product is dissolved in the hot solvent the insoluble
impurities (type 1 above) can be removed by hot filtration (p. 98). When the hot
solution is allowed to cool, the solution becomes saturated with the desired
compound and it precipitates from the cold solution. The cold solution does
not become saturated with the lower concentration of the contaminants of type
2, which therefore remain in solution. Coloured impurities (type 3) can be
removed by absorption using charcoal as described on p. 96.
The recrystallization process can be divided into three separate steps:
1. Selection of a suitable solvent.
2. Recrystallization of the crude compound.
3. Drying the purified solid.
Solvents for recrystallization

You can decide on a suitable solvent for recrystallization of your crude
chemical product in several ways:
• The experimental protocol may tell you which solvent to use.
• If you know the identity of the compound you have made, reference texts
may indicate a suitable solvent (e.g. Lide, 2000; Buckingham and
Macdonald, 1995).
• If you are not sure of the identity of the compound you have prepared or
if it is a new compound, you must carry out a 'solvent selection' to find
out which solvent is the most appropriate (p. 93).
When selecting a suitable solvent for recrystallization, chemists work to the

general rule 'like dissolves like' when considering the polarity of the chemical to
be recrystallized and the polarity of the solvent. In general solvents for
recrystallization are classified in terms of polarity and miscibility. Solvent
polarity depends upon the overall distortion of the electron clouds in the
covalent bonds within the solvent molecules, resulting in a dipole. The greater
Recrystallization
the dipole, the greater the polarity of the solvent, e.g. trichloromethane has
three polarized C-C1 bonds, giving an overall dipole to the molecule and
producing a good solvent for many molecules, but in tetrachloromethane all of
the C-C1 dipoles cancel out, giving a non-polar solvent with different solvent
properties in comparison. An additional feature that adds solvating power is
hydrogen bonding. Thus water, a polarized molecule, can form hydrogen bonds
with oxygen and nitrogen atoms in solutes, dissolving them efficiently.
When you are choosing a solvent for recrystallization, look for the
following general characteristics:
• A high dissolving power for the solute at high temperature and a low
dissolving power at room temperature or below, so that a high recovery
of purified compound can be achieved.
• A high or negligible dissolving power for the impurities, so that they will
either be filtered off or remain in solution.
• A relatively low boiling point, to facilitate drying the purified compound.
The miscibility of solvents with each other must be taken into account
when attempting mixed-solvent recrystallizations (see p. 95); the properties of
some common recrystallization solvents, which you are likely to encounter in
your laboratory work, are shown in Table 13.1. Remember that this is only a
general list of solvents; further information on solvent properties can be
found in standard textbooks such as Harwood et al. (2000, p. 133),
Loewenthal (1990, p. 146) and Furniss et al. (1989, p. 137).
Table 13.1 Selected solvent properties
Water 100 V. high Yes Always use when suitable:

salts, aromatic acids
Ethanol 78 High Yes Flammable: alcohols, acids,
amides
Propanone 56 High Yes Flammable: carbonyl
compounds
Ethyl ethanoate 78 Medium No Flammable: esters
Dichloromethane 41 Medium No Toxic: halogen compounds,
ethers
Toluene 110 Low No Flammable: hydrocarbons
Light petroleum 40-60 V.low No Flammable: hydrocarbons
Solvent selection
To find a suitable solvent for recrystallization you must carry out a series of
tests measuring the solubility of your crude compound in a series of solvents
(cold and hot) of varying solvent polarity. This series of tests is called 'solvent
selection' and is carried out on a test tube scale but, as your technique
improves, you can carry out the tests using semi-micro scale since you will
use less of your compound during the process. The procedure for solvent
selection at the test tube scale is described in Box 13.1 and modification of
the procedure to semi-micro scale requires only a corresponding reduction of
the quantities of solvent and solute used.
~ When carrying out the solvent selection experiments

you must always cool the solution after heating. Do not assume that if
the compound is insoluble in the cold solvent and dissolves when
heated, it will always precipitate on cooling.
Recrystallization
Recrysta 11ization
Mixed solvents
When no single solvent is found to be suitable for recrystallization, then a
mixed solvent system must be used. There are three essential properties
required for a pair of solvents to be used in a mixed-solvent system:
I. The two solvents must be miscible in all proportions over the
temperature range to be used.
2. The solute must be insoluble in one of the solvents.
3. The solute must be soluble in the other solvent.
It is an advantage if the two solvents have similar boiling points within 20-
30°C.
Suitable common solvent pairs from the solvents given in Table 13.1 are
water/ethanol and dichloromethane/light petroleum (b.pt. 40-60°C), but
many other combinations are possible. One of the most frequently
encountered mixed solvent systems is 'aqueous ethanol' (water/ethanol) in
which the compound to be recrystallized is insoluble in water and very
soluble in ethanol.
You can identify the solubility characteristics of your compound using the
solvent selection procedure shown in Box 13.1 and modifications for mixed
solvent selection are given in Box 13.2.
The recrystallization process

Having chosen a suitable solvent system, the process to be used to purify the
bulk of your impure compound can be separated into several distinct steps:
Recrystallization
• Dissolution of the solid using a single-solvent or a mixed-solvent system.

• Decolorization using charcoal: even if your compound is white,
decolorization will improve its appearance significantly.
• Hot filtration (p. 98) to remove solvent-insoluble impurities and
charcoal.
• Cooling, to produce the crystals.
• Collection of the crystals by suction filtration (p. 28).
• Drying (p. 39).
It is important to remember that for a successful recrystallization, you need
to use equipment of a size appropriate to the amount of solid and the volume
of solvent you are likely to use. You can estimate the volume of solvent to be
used by extrapolation of the data from your solvent selection tests. In general
terms, conical flasks, beakers and round-bottom flasks should never be more
than half-full of solution but, on the other hand, using small volumes of
solutions in large flasks will result in losses of the compound on the sides of
the vessels.
Dissolution
To carry out a single-solvent recrystallization (Box 13.3) you must get the
compound into solution and this is achieved by suspending it in the
appropriate cold solvent, found in the solvent selection process, and then
heating the mixture to dissolve the solid. The equipment used will depend on
the boiling point of the solvent, its flammability and toxicity. Some general
systems are shown in Table 13.2.
Table 13.2 Solvents for recrystallization
Water 100 Conical flasks, Burner, hot plate None

beakers
Ethanol 78 Conical flasks Water bath Fume cupboard
Propanone 56 Conical flasks Water bath Fume cupboard
Ethyl ethanoate 78 Conical flasks Water bath Fume cupboard
Dichloromethane 41 Reflux Water bath Fume cupboard
Toluene 110 Reflux Hot plate, mantle Fume cupboard
Light petroleum 40-60 Reflux Water bath Fume cupboard
When heating solvents in conical flasks and beakers you should cover the
top of the flask with a clock-glass or watch-glass to prevent excessive
evaporation of the solvent, resulting in the formation of crystals on the sides
of the flask above the surface of the solution. Do not forget to take the
appropriate 'anti-bumping' precautions (p. 31), because you may need to boil
the mixture for several minutes to achieve complete dissolution of the
chemical(s). You should use a glass rod, a wooden 'boiling stick' or anti-
bumping chips in conical flasks of volume up to 500mL; magnetic 'fleas' or
anti-bumping granules in round-bottom flasks in reflux apparatus (p. 116);
magnetic 'fleas' or stirring bars in large-volume glassware (> 500 mL).
Decolourization
Small amounts of coloured impurities can be removed from your product by
absorption on finely divided charcoal, which is then removed in the hot
filtration process (p. 98). In general, you should use charcoal in every
Recrystallization
recrystallization since the colour of white and coloured compounds is

improved by 'decolorization'.
The amount of charcoal used should be about 2% by weight of the sample
to be recrystallized. Note the following important points:
• Add the charcoal only when the solution has been formed. If you add
charcoal to the cold solution, you will not be able to see when all of the
compound has dissolved.
• Do not add charcoal to a boiling or a very hot solution otherwise the
solution will boil extremely vigorously, usually boiling out of the flask or
reflux apparatus.
• To add charcoal to a hot solution, remove the flask from the heat source
and then let it cool a little (not enough to cause precipitation). Add the
charcoal either from a spatula (into a conical flask or beaker), a paper
funnel (p. 25) or powder funnel (into a round-bottom flask with a
ground-glass joint in reflux apparatus), and then reheat the solution.
Recrystallization
Hot filtration
This is a modification of gravity filtration (p. 28), designed to remove
solvent-insoluble impurities, charcoal, anti-bumping granules or magnetic
'fleas' from the hot solution before cooling the solution to form the crystals
of purified product.
~ Hot filtration is used to prevent cooling of the solu-

tion during the filtration process, which would result in the formation of
crystals in the filter paper.
For a successful hot filtration the solution must pass through the filter paper
and filter funnel into the collection flask as quickly as possible so that cooling
and crystallization do not occur. The following points should be noted:
• Always use a fluted filter paper (p. 28).

• Always use a 'stemless' glass filter funnel because cooling and thus
crystallization may occur in the funnel stem causing a blockage.
• Always heat the filter funnel and filter paper, either by pre-heating them
in an oven or by using boiling solvent in the collecting flask (Fig. 13.1).
• If filtration is rapid, keep the filter cone 'topped up' with the hot solution
stemless funnel being filtered, since this keeps the filter cone and filter funnel hot. If you
allow the filter cone to empty of liquid, cooling and crystallization may
occur.
• When attempting a hot filtration with a mixed-solvent system, always
hotplate
filtrate refluxing (or water bath) ensure that the 'ideal' solvent ratio has not been reached, i.e. there is not
gently
enough of the poor solvent to cause immediate precipitation as a result of
slight cooling during the hot filtration. The solvent ratio can be adjusted
to the 'ideal' ratio for maximum recovery, after filtration and before
cooling (Box 13.4).
Fig. 13.1 Filtration of a hot solution.
Cooling
Rapid cooling in an ice-water bath ('crash-crystallization') usually produces
small crystals occluded with mother-liquor, whereas slow cooling by allowing
the collection flask to stand on the laboratory bench often produces large
well-defined crystals. Remember to:
• cover the top of the collection flask with a watch- or clock-glass to
prevent solvent evaporation and entry of dust into the flask: do not use
rubber bungs or corks since they may be pulled into the flask as it cools
(p. 21);
• clamp the flask in place, if you use an ice-water bath, otherwise it may
fall over as the ice melts and the volume of water increases;
• make sure that the solution is cold, even after slow cooling, so that
maximum precipitation of the solid occurs.
If no crystals appear on cooling, you will have formed a supersaturated
solution and, to induce precipitation of the solute, you must provide sites for
nucleation and crystal growth. This can be achieved by either seeding the
solution by adding a few crystals ('dust') of the crude compound or
scratching the inside of the flask at the surface of the liquid, using a Pyrex@
glass rod (Fig. 13.2).
Collection of the crystals

You should collect the purified crystals by suction filtration by the procedure
described in Box 5.2 (p. 30). Remember to transfer the crystals from the
Recrystallization
collecting flask to the filter using a little of the filtrate: do not use fresh
solvent. The filtrate is a saturated solution of the compound being
recrystallized and cannot dissolve any more solute, but fresh solvent will
dissolve some of your product resulting in an inefficient recrystallization
process.
Recrystallization
(bl
Fig. 13.2 'Scratching': (a) right; (b) right; (c) wrong.
Drying
The purified compound should be dried by the appropriate method as
described on p. 39. If you do not know the melting point of your
compound, you should always carry out a test by placing a small amount
on a watch-glass in the oven, before committing the bulk of your
chemical.
Detailed procedures for single-solvent and mixed-solvent recrystallizations
are shown in Box 13.3 and Box 13.4 respectively and the modifications
necessary for the use of other solvent systems can be worked out from the
information in Table 13.2.
Problems in recrystallization
There are three common problems encountered during recrystallization:
1. The compound crystallizes in the filter funnel during hot filtration. This
is because the solubility of the solute decreases rapidly with temperature
and the slight cooling during hot filtration causes precipitation of the
solid, even though you are heating the funnel. The answer is to use more
than the minimum. amount of solvent and then evaporate off the excess
before cooling.
2. The compound does not recrystallize. There are two reasons: you have
used too much solvent and you must evaporate off some solvent before
cooling, or you have formed a supersaturated solution and you must
'seed' or 'scratch' the solution (see p. 98).

Recrystallization
3. The compound precipitates as an oil. This is because compounds with

low melting points often come out of a concentrated solution above their
melting point. In such cases more solvent should be added and the
compound redissolved and cooled so that precipitation is retarded to the
temperature at which the crystalline solid comes out of solution. Often
'scratching' the hot solution as it cools can prevent 'oiling out'.

------~G Solvent extraction
This technique separates the components of chemical mixtures by using the

dissimilar solubility properties of the components of the mixture in
different solvents. Extraction is used mainly to purify a reaction product
partially before final purification by recrystallization (p. 92) or distillation
(p. 107). The two common types of extraction process used in the labora-
toryare:
I. Liquid-liquid extraction: this uses two immiscible solvents; the desired
compound in solution or suspension in one solvent is extracted into the
other solvent. For example, covalent organic compounds are extracted
from aqueous solution into dichloromethane, leaving the ionic by-
products or reagents in the aqueous phase.
2. Solid-liquid extraction: this involves the use of a solvent to remove
solvent-soluble components of a solid mixture.
Liquid-liquid extraction
Several experimental processes in practical chemistry are based on liquid-
liquid extraction:
• 'Extraction': where a solid or liquid suspended or dissolved in one solvent

is extracted into another. This technique can be used to separate covalent
molecules from ionic compounds in an aqueous solution or suspension.
• 'Washing': where ionic species are removed from a non-polar solvent by
extraction into water.
• 'Acid-base extraction': where covalent molecules are converted into their
salts and thus removed from a non-polar solvent into water, while neutral
covalent species will remain in the non-polar solvent, as shown in
Table 14.1.
Table 14.1 Examples of acid-base extraction chemistry
NaOH
ArCOOH + RH ArCOO- Na" + RH
CH;CJ;
Acid Neutral Salt Neutral
insoluble in H20 insoluble in H20 soluble in H20 insoluble in H20
soluble in CH2CI2 soluble in CH2CI2 soluble CH2CI2
HCI
ArNH2 + RH CH;CJ; ArNHt CI- + RH
Amine Neutral Salt Neutral
soluble in CH2CI2 soluble in CH2CI2 soluble CH2CI2
ArCOOH + ArOH ArCOO- Na" + ArOH
Acid Weak acid Salt Weak acid

soluble in CH2CI2 soluble in CH2CI2 soluble CH2CIz
All of these processes involve mixing the two immiscible solvents, one of
which contains the mixture, in a separatory funnel (p. 103) and shaking the
funnel to promote the extraction process. The immiscible layers are allowed
to reform and are then separated.

Solvent extraction
For liquid-liquid extraction, water is usually the polar solvent. Since

most extractions involve getting the required compound into the organic
solvent (or removing unwanted ionic chemicals from it), it should have
good solvent power for the desired compound and a low boiling point for
ease of removal and recovery of the compound. The common organic
solvents used in liquid-liquid extraction are diethyl ether (ethoxyethane)
b.pt. 34 QC, dichloromethane (DCM) b.pt. 41 QC and ethyl acetate (ethyl
ethanoate) b.pt. 77 QC. Dichloromethane is denser than water and forms
the lower layer, whereas diethyl ether and ethyl acetate float on water and
are the upper layer.
Partition coefficients
The theory of liquid-liquid extraction is based on the equilibrium between
the concentrations of dissolved component in the two immiscible liquids,
when they are in contact. The equilibrium constant for this process is called
the partition coefficient or distribution coefficient and is given by:
concentration of solute in liquid I
[14.1]
K = concentration of solute in liquid 2
You only need to calculate such quantities if:
• you are carrying out specific experiments to determine partition

coefficients, when you will be given specific instructions or references to
the appropriate literature;
• the solute has appreciable solubility in both solvents.
The reason calculation is not necessary is that, in the overwhelming
majority of extractions you will carry out, the conditions used are designed
to ensure that the components will be almost totally soluble in one of the
liquids and almost insoluble in the other, since complete separation is
required. The number of extractions needed to extract a water-soluble solute
into an immiscible organic phase can be calculated from the following
relationship:
Kv )11
WI1 = Wo ( Kv +s [14.2]
where K = partition coefficient of the solute, v = volume (mL) of aqueous

solution of the solute, s = volume (mL) of immiscible organic solvent, Wo =
weight of solute initially in the aqueous layer, WI1 = weight of solute
remaining in the aqueous layer after n extractions, and n = number of
extractions. Evaluation of this expression shows that, for a fixed volume of
solvent, it is more efficient to carry out many small extractions than one big
one.
Separatory funnels
These come in a range of sizes from 5 mL to 5000 mL and there are two
general types: parallel sided and cone shaped (Fig. 14.1). Cone-shaped
separatory funnels are made of thin glass and should be supported in a ring
(p. 104). Small-volume cone-shaped funnels « 100mL capacity) and
parallel-sided separatory funnels should be clamped at the ground-glass joint
at the neck (p. 27).
Separatory funnels will have glass or Teflon@ taps with a rubber ring and
clip or screw cap on the end to prevent the tap slipping from the barrel, or a

Solvent extraction
Rotaflo'P tap. You must ensure that the tap assembly is in good condition by
making the following checks before starting work:
• For glass taps: disassemble the tap by first removing the clip and ring or
cap from the tap (note the order of the component parts for
reassembling). Dry the tap and barrel with tissue, add a light smear of
grease to the tap (making sure you do not clog the hole in the tap) and
reassemble the tap and fittings, turning the tap to ensure free movement.
Support the separatory funnel in position and add some of the organic
solvent to be used (2-3 mL) to the funnel, with the tap closed, to check
that the tap does not leak. If the tap leaks, disassemble and regrease.
• For Teflon® taps: disassemble the tap, wipe the tap and barrel with clean
tissue, reassemble without grease, check for free movement of the tap and
for leakage as described above. When you have finished using the funnel,
loosen the clip/cap on the tap since Teflon® will flow under pressure and
the tap may 'seize' in the barrel.
• For a Rotaflo® tap: unscrew the tap from the funnel and ensure that
the plastic locking thread is in place (Fig. 14.2). If it is not present,
consult your instructor and obtain a replacement. Dry the barrel of
the tap and the tap with a tissue and reassemble. Do not grease the
Rotaflo® tap.
The general procedure for using a separatory funnel for extraction is

described in Box 14.1 and there are five additional practical tips to aid your
success:
1. Label all flasks to avoid confusion.

2. Never throwaway any of the separated liquids until you are absolutely sure
of their identity.
3. Always transfer solvents into the separatory funnel using a stemmed
filter funnel so that solids and liquids will not stick to the inside of the
Fig. 14.1 Separatory funnels. joint and prevent a good seal when you insert the stopper and then
invert the funnel.
4. Always place a 'safety' beaker under the separatory funnel to collect
liquid just in case the tap leaks (Fig. 14.3a).
funnel inverted stopper
separated liquid layers

+-- covered metal ring
(e.g. PVC)
plastic locking ring stand stopcock (closed!)
I ~ glass thread
beaker
)
Rotaflo® tap
stopper
(ready for use)
(a) (b)
Fig. 14.2 A Rotaflo® tap. Fig. 14.3 A separatory funnel (a) ready to use and (b) in use.

Solvent extraction
5. Always take the stopper from the separatory funnel before you attempt
to allow liquid to run from the funnel. If you do not remove the stopper
from the top of the funnel, a vacuum is formed in the funnel after a little
of the liquid has run out. Air will be sucked into the funnel through the
outlet stem causing bubbles, which will remix your separated layers. If
your funnel is equipped with a Quickfit'f stopper, it is good practice to
take the stopper out of the top and put it back upside down (Fig. l4.3b).
This ensures that no vacuum is formed and that organic vapours do not
Fig. 14.4 Holding a separatory funnel. escape easily from the flask.
Solid-liquid extraction
In this process the components of a solid mixture are extracted into a solvent.
The 'batch process', analogous to liquid-liquid extraction, involves grinding
the solid to a fine powder, mixing it with the appropriate solvent, and
filtering off the solid by gravity (p. 27) or under vacuum (p. 30) and then
evaporating the solvent (p. 121) from the extract solution. However, a more

Solvent extraction
elegant 'continuous extraction process', called Soxhlet extraction, is available

condenser
when the most appropriate solvent is known.
The apparatus for Soxhlet extraction is shown in Fig. 14.5 and comprises
a flask containing the solvent, a Soxhlet extractor and a reflux condenser
(p. 116). The solid to be extracted is placed in a porous thimble, made from
clamp
hardened filter paper, and the solvent is heated so that its vapour flows past
paper
the thimble, condenses and fills the extractor with hot solvent to extract the
thimble
solid to be Soxhlet
solid. When the extractor is full, the solvent (together with the extracted
extracted apparatus material) siphons back into the solvent flask and the process is repeated
automatically. The advantage of this procedure is that fresh solvent
continually extracts the solid, which is concentrated in the flask. The
disadvantage is that the compound extracted is kept at the boiling point of
clamp the solvent for a prolonged period. Soxhlet extractors come in sizes of
10 mL to 5000 mL, based on the volume of solvent contained in the
extractor. The procedure for using a Soxhlet extraction system is described
boiling in Box 14.2.
stones
heat source
(mantle, water ---+-
bath, etc.]
Fig. 14.5 A Soxhlet extraction system.

-------0 Distillation
Distillation is used to separate the components of a liquid mixture by

vaporizing the liquids, condensing the vapours and collecting the liquid
condensate. Separation is the result of the differing boiling points of the
individual constituents of the mixture and the efficiency of the distillation
column. You may be required to purify a liquid by distillation, which involves
the removal of small quantities of impurities, or to separate a mixture of
liquids by Factional distillation, each of which can then be purified by
distillation.
You will meet several types of distillation process each applicable to
different situations depending on the chemicals to be purified or
separated.
• Simple distillation: used for separating liquids, boiling below 200 QC at
atmospheric pressure, from other compounds. For effective separation
there should be a difference in the boiling points of the components of at
least 80 QC.
• Fractional distillation: used for separating components of liquid mixtures,
which have boiling points differing by more than 25 QC, at temperatures
below 200°C.
• Vacuum or reduced-pressure distillation: used for separating liquids boiling
above 200 QC, when decomposition may occur at the high temperature.
The effect of distilling at reduced pressure is to lower the boiling point of
a liquid. This technique can be applied to both simple distillation and
fractional distillation.
• Steam distillation: used for separating mixtures of chemicals such as oils,
resins, hydrocarbons, etc., which are essentially insoluble in water and
may decompose at their boiling points. Heating the compounds with
steam makes them distil below 100 QC.
Distillation equipment
Apparatus used for the various types of distillation has several general
features:
• Distillation flask: usually round bottom or pear shaped with one or two
necks (to allow a vacuum bleed to be fitted).
• Still-head: to hold the thermometer and to channel the vapour into the
condenser. For fractional distillation, the fractionating column is fitted
between the distillation flask and the still-head.
• Condenser: usually with circulating cold water.
• Take-off adapter: to allow the distillate to run into the collecting vessel.
For vacuum distillation, the adapter will have a vacuum inlet tube and
could have three 'arms' to allow three fractions to be collected without
breaking the vacuum.
• Receiving (collection) vessel: this can be a test tube, a measuring cylinder,
a conical flask, or a round-bottom flask with a ground-glass joint.
No matter what type of distillation you are attempting, it is essential that you
assemble the apparatus correctly, since you are dealing with hot, often
flammable, liquids and vapours. A typical simple distillation apparatus is
shown in Fig. 15.1 and the method of assembly is described in Box 15.1.

Distillation
~ You must ensure that all the ground-glass joints are

seated properly and the apparatus is clamped firmly so that no move-
ment of the joints will allow vapours to escape.
screwcap adapter Simple distillation

with thermometer
The procedure for doing a simple distillation is described in Box 15.2 and
joint clips
you should note the following points:
• Do not distil to dryness, i.e. no liquid remaining in the distillation flask,

since this causes overheating and charring (decomposition) of the residue.
Always leave I or 2 mL of liquid in the distillation flask.
• Initially, some liquid may distil while the temperature rises rapidly. This is
termed 'forerun', for example often a small amount of solvent from an
extraction process (p. 102): collect and then, when the temperature
stabilizes, collect the desired compound in a clean receiving vessel.
• Towards the end of the distillation, the temperature may begin to rise
again: this will be the higher boiling impurity. Stop heating, quickly
remove the receiving flask containing your pure compound and collect
Fig. 15.1 Apparatus for simple distillation.
these last few drops, termed 'tailings', in a fresh receiving vessel.
Fractional distillation
Simple distillation is not very effective in separating liquids unless their
boiling points are at least 80 QCapart and a better separation can be achieved
if a fractionating column is used. There are many types available (Fig. 15.2)
and the device brings the ascending vapours into contact with the condensing
(in the fractionating column) liquid and amounts to a succession of many
simple distillations in which the descending liquid strips the high-boiling
component from the ascending vapour. Overall, the lower boiling component
distils first and cleanly. The column packing, usually glass beads, helices or
'fingers', gives a large surface area for contact of the ascending vapours and
descending liquid. After the first fraction has distilled, the temperature will
rise and the rate of distillation will slow. This is an intermediate fraction
containing a little of both components of the mixture. Finally, the
temperature will become constant and the pure higher boiling compound will
distil. The procedure for fractional distillation is given in Box 15.3.
open column Vigreux column Glaisen flask Glaisen-Vigreux flask

Fig. 15.2 Common fractionating columns.
~ A successful fractional distillation relies on thermal

equilibration of the components in the fractionating column. You must
allow the column to be heated by the vapours of the mixture.

Distillation

Distillation
Vacuum or reduced-pressure distillation

Distillation at reduced pressure is used to distil liquids with few impurities or
to fractionate the components of liquid mixtures with high boiling points,
which would decompose if distilled at atmospheric pressure. The
modifications to the distillation apparatus (Fig. 15.1) required for reduced-
pressure distillation are:
• A vacuum source: this can be a water pump (see p. 29) with an 'anti-suck-
back' trap producing a vacuum of 15-25 mm Hg, at best; or a rotary vacuum
pump (consult your instructor about its use since it is an expensive piece of
equipment and easily contaminated or damaged), which will evacuate down
to 0.1 mm Hg. Two-stage dry vacuum pumps produce a vacuum of 1~5 mm
Hg, are resistant to organic and acid vapours and are easy to use.
• A manometer to measure the pressure (vacuum) in the apparatus, since the
boiling point of a liquid varies with pressure. Two types are in common use
(Fig. 15.3): the Anschutz manometer, which gives a constant reading of the
vacuum throughout the distillation, and the Vacustat'P, which is used to
take a 'sample' of the vacuum at a given instant. Vacustats'P are very
accurate and are more often used in combination with a rotary oil pump.
• A take-off adapter, which permits the collection of several fractions
without stopping the distillation to change the receiving flasks. A
receiving adapter, termed a 'pig' (Fig. 15.4), can be rotated on the end of
the condenser to collect three fractions.
• Appropriate 'anti-bumping' measures: reduced-pressure distillations are
notoriously prone to 'bumping' (p. 31) and anti-bumping granules are
ineffective. You can use a magnetic 'flea' in conjunction with a hot plate-
stirrer and oil bath; an air bleed, made by pulling out a Pasteur pipette
into a fine capillary using a microburner (consult your instructor),
inserting it into a screwcap adapter (p. 43) and placing it in the second
neck of the distilling flask. The vacuum pulls a fine stream of air into the

Distillation
left-hand limb filled -"'-11-1._

with mercury
-
to aspirator
to apparatus
-
Anschutz manometer
via trap
Fig. 15.3 Manometers for vacuum distillation.
vacuum
'pig' type
receiving
adapter
receiving
flask
Fig. 15.4 'Pig'-type receiving flask adapter.
flask and forms small bubbles which agitate the liquid during distillation;
or a wooden boiling stick for small-scale distillations of short duration,
since the vacuum will pull air from the stick and agitate the liquid.
• A three-way tap inserted between the vacuum source and the manometer,
so that you can control the vacuum applied to the distillation system.
A typical system for reduced-pressure distillation is shown in Fig. 15.5 and

the procedure, using a water pump, is described in Box 15.4.
Quickfit® thermometer
water pump 'trap'
air bleed manometer

with clip
Fig. 15.5 Apparatus for vacuum distillation.
Kugelrohr distillation
This is also known as short-path distillation or bulb-to-bulb distillation. It is a
procedure for reduced-pressure distillation, which almost eliminates losses
owing to the size of the apparatus, and is particularly useful for small volumes of
liquids. The liquid sample is placed in a small round-bottom flask, connected to
a series of bulbs (Fig. 15.6) and then heated and rotated (to prevent bumping)

Distillation
under vacuum in an electric oven. The liquid distils from the flask inside the
oven to a cold bulb outside the oven. Since the distillation path is very short the
length of a ground-glass joint losses are minimized. The use of several bulbs as
receiver flasks allows a small volume of mixture to be fractionated by varying
the temperature in the oven. The distillates can be removed from the bulbs by
either washing into another flask with a low-boiling-point solvent or using a
'bent' Pasteur pipette (Fig. 15.7) since the bulbs can only be held horizontally.
oven motor
tapered joints vacuum
heating control
temperature gauge
Fig. 15.6 Kugelrohr distillation apparatus.
Steam distillation
This process can be used to separate water-insoluble covalent compounds
from crude reaction mixtures or to isolate volatile natural products, e.g.
terpenes from plant tissue. The crude mixture is heated with water and steam
and the steam-volatile material eo-distils with the steam and is then
Fig. 15.7 A 'bent' Pasteur pipette.
condensed with the water and collected. You will then need to carry out an
extraction (p. 102) to separate the water and the insoluble component.
400~
°C
700::
600::
300~
500::
400:
200~
300:
200:
Fig. 15.8 Nomograph for estimating boiling 100~

point at reduced pressure. To use it, draw a line pressure in mm Hg
b.pt. corrected
between the recorded pressure and the boiling to 760 mm Hg
point at 760 mm Hg, and then extend the line to
the observed boiling point scale. This point is
the boiling point at the reduced pressure. In
this example, a liquid b.pt. of 250 QCat 760 mm
Hg will boil at 118°C at 10 mm Hg. observed b.pt.

Distillation

Distillation
water for replenishing

distillation pot
clamp
Claisen adapter
clamp
heat source
Fig. 15.9 Apparatus for steam distillation.
The steam required for a steam distillation can be provided by an external

source, such as piped steam in the laboratory or steam generated by heating
water in a flask, which is then piped into the distilling flask. Steam is very
dangerous and the safest way to generate steam is to heat the compound with
a vast excess of water in the distilling flask: steam is generated in situ. The
equipment for steam distillation is illustrated in Fig. 15.9 and the procedure
is described in Box 15.5.
~ You must never attempt a distillation in a completely

closed system. Always check that the expanding vapours can escape.

Distillation

-----~G _R_ef_lu_x _
Reflux is one of the most common techniques you will encounter in your
chemistry laboratory classes. Since many reactions between covalent
compounds are slow processes rather than instantaneous reactions, prolonged
heating forces the equilibrium to give an acceptable amount of product. In the
reflux process, the reactants are dissolved or suspended in a suitable solvent, the
solvent is boiled and then condensed so that it returns to the reaction flask.
Once set up, a reaction carried out under reflux can be run for minutes, hours
or even days to promote the required reaction. The basic components for a
reflux apparatus are:
• a reaction flask;
• a reflux condenser;
• a heat source (see p. 32);
• a coolant source, usually water, for the condenser.
The procedure for setting up a simple reflux apparatus (Fig. 16.1) is given
in Box 16.1.
Reaction flasks
Round-bottom or pear-shaped reaction flasks are preferred, but note that
stirring with the usual type of magnetic 'flea' is not possible in pear-shaped
flasks. The flasks can have multiple necks so that the apparatus can be
configured for temperature measurement, addition of solids or liquids,
mechanical stirring and inert atmosphere work (p. 125). No matter which
arrangement of components is used, always clamp the reaction flask at the
neck and keep the heaviest components (such as an addition funnel
containing another chemical) vertically above the flask. A condenser will still
function at 30° from vertical and it is not very heavy.
Condensers
For general-purpose work, these are usually single-surface or double-surface
types (Fig. 16.2): the double-surface condenser is used for low-boiling point
solvents such as dichloromethane, diethyl ether or light petroleum (b.pt. 40-
60 QC).
condenser
heating bath
(oil etc.)
magnetic stirrer/
hot plate
1
_
"'"
~ ©~
~=~----~~~ single double
coil
surface surface
Fig. 16.1 Apparatus for simple reflux. Fig. 16.2 Condensers.

Reflux
outer rubber gasket Rubber tubing for coolant water is attached to the condenser in two ways:
-
tOCOOlant~
pipe ~.
.-
"=>=
1. If the condenser has glass inlet and outlet pipes with a 'knuckle', wet the
inside of the rubber tube with a little water and slide it onto the pipe and
past the 'knuckle'. The rubber tubing must be a tight fit otherwise it may
slide off over a period of time.
2. Modern condensers have plastic adapters, which can be attached to the
tubing and then screwed on the threaded inlet and outlet pipes. Slide the
rubber tubing onto the moistened 'pipe' on the adapter, and then screw
inner rubber gasket the adapter onto the condenser. You must ensure that the adapter has a
rubber gasket on the inside of the threaded portion (Fig. 16.3), otherwise
Fig. 16.3 Plastic connectors for condensers.
it will leak water at the condenser inlet or outlet.
~ You should always attach rubber tubing to a conden-

ser before fitting it to the apparatus.
Drying tubes
Water can get into your reaction by condensation from the atmosphere or
by condensation of the steam produced in a water bath. To exclude
water, you should fit a guard tube containing a solid drying agent such as
anhydrous calcium chloride or calcium sulphate to the top of the
condenser. A typical guard tube is shown in Fig. 16.4; use a coarse-sized
drying agent rather than a fine powder, which would 'cake' very quickly
as it absorbs moisture.

Reflux
Reflux with addition of chemicals

Instead of stopping the reaction and opening the apparatus, you can put in
the new chemicals using an addition or 'dropping' funnel. Addition funnels
are separatory funnels (see p. 119) with a ground-glass joint on the stem, and
there are two types (Fig. 16.5):
1. Addition funnels: when you add a liquid or solution from the funnel
CaCI21umps to the reaction flask, you must take out the stopper, otherwise a
vacuum is formed and the liquid does not flow out (see p. 105). This
Fig. 16.4 A CaCI2 guard tube.
is a disadvantage when using compounds with irritating or toxic
vapours or which are air sensitive. The simplest solution to this
problem is to fit a guard tube, instead of a stopper, to the addition
funnel and then the liquid or solution will flow easily into the
reaction flask.
2. Pressure-equalizing dropping funnels: these have a side arm linking
the reservoir of the funnel to the inlet stem below the tap. The
pressures in the reservoir and the reaction flask are equal, and liquid
will flow into the reaction flask even when the stopper is in place in
the funnel. Pressure-equalizing dropping funnels are very expensive
and are normally only used for inert atmosphere reactions (p. 119).

Reflux
The addition of dry solids to refluxing reactions requires special equipment

and you should refer to the appropriate texts (Errington, 1997, p. 125;
Harwood, et al., 2000, p. 88; Furniss, et aI., 1989, p. 82). The simplest
approach is to dissolve the solid in a small amount of the solvent being used
and add it as a solution, using an addition funnel.
Reflux with mechanical stirring

When a magnetic stirrer is unsuitable, e.g. III reactions involving viscous
liquids or mixtures of solids and liquids, a mechanical stirrer must be used. A
mechanical or overhead stirrer system comprises:
1. An electric variable speed motor connected to the mains supply.
2. A flexible connector, usually a short length of rubber tubing.
guard tube 1
o
clamp
clamp
condenser
Claisen
adapter
two-neck flask
magnetic
stirrer bars
(a) (b)
_"\ I, "'",
:'@/~~ ::'@/~ _,\ I1 -.
Fig. 16.5 Adding chemicals to a reflux

apparatus: (a) addition funnel; (b) pressure-
-@-
~-=,-----~=~
J magnetic stlrrer/I
hotplate _
~=~-----~~
-@-
equalizing funnel.
(a) (b)
sealed stirrer
guide
clamp
three-neck flask
heat source
(bath, heating mantle)
(c)
Fig. 16.6 Apparatus arrangements for reflux with addition: (a) heating a reaction
mixture to reflux during the addition of a reagent; (b) heating a stirred reaction
mixture during the addition of a reagent; (c) heating and overhead stirring of a
reaction mixture during the addition of a reagent.

Reflux
3. A stirrer gland, which fits into the top ground-glass joint of the flask
acting as a seal for the refluxing vapour and a guide for the stirrer shaft.
There are several types of gland available, but the best is now made from
Teflon@ and needs no lubrication. If this is not available, a screwcap
adapter in which the rubber sealing ring has been lubricated with a touch
of silicone oil can be used. Note that the stirrer shaft should rotate in the
adapter, not the adapter in the joint of the flask, so do not tighten the
screwcap too much.
4. A Teflon® paddle which swivels on the end of the stirrer shaft so that it
can pass through the ground-glass joint on the top of the flask.
Setting up the apparatus for reflux with mechanical stirring is a precision

task and is described in Box 16.2. The major problems encountered are:
• The weight of the stirrer motor high up on the support stand: use a large
support stand with a heavy base or use a support framework, which is
screwed to the laboratory bench. Besides the motor's weight, the torque
of the motor can cause 'whipping' in the support stand.
• The motor, stirrer gland, stirrer shaft and reaction flask must be
absolutely vertical and concentric, otherwise the glass stirrer shaft will
snap.
• Since there will always be a little sideways movement when the stirrer is
operating, the flask and the motor should be clamped in position on the
same stand. Condensers and addition funnels should not be clamped.
A selection of configurations, suitable for most reflux procedures is shown in
Fig. 16.6.

------~0
Evaporation
Evaporation is the process in which the solvent of a solution is converted into

a vapour to leave a solid or liquid solute. There are many applications of
evaporation, ranging from the slow evaporation of water from a solution of
an ionic compound to leave hydrated crystals, e.g. crystallization of
CuS04.5H20, to the evaporation of large volumes of low-boiling-point
solvent under reduced pressure in the extraction of an organic compound.
Since crystallization by evaporation is a specific technique for a relatively
small range of compounds, the term evaporation is generally interpreted as
the removal of the solvent from a solution.
Evaporation of solvents
There are three commonly used techniques for solvent evaporation:
I. Evaporation from open flasks on a steam bath.
2. Rotary film evaporation.
3. Gas 'blow-down'.
All these techniques have advantages and disadvantages. Where your
experimental protocol may simply state 'the solvent is evaporated off, you
should select the most appropriate procedure based on:
• the volume of solvent to be removed;

• the amount of solute in solution;
• the relative boiling points of the solvent and solute;
• the next step in the experimental procedure.
Evaporation from open flasks

This is useful for evaporating small volumes (~ 25 mL) of low-bailing-point
solvents « 70 QC) from solutions containing a solute which has a boiling
point above 110QC.Place the solution in a beaker or conical flask, containing
a glass rod or boiling stick, onto a steam bath (maximum temperature
achievable is 100QC) in a fume cupboard and evaporate the solvent until
boiling ceases. The obvious advantage is simplicity; the disadvantages include
environmental concerns of release of solvents in the atmosphere,
contamination of the solvent by condensed steam and incomplete solvent
removal due to the 'hold-up' volume of the flask.
Rotary film evaporation

This method, which is also known as rotary evaporation or 'rovap', is the
technique of choice for the removal of large volumes of volatile solvents
from solutions, e.g. from extractions and column chromatography (p.
217). Rotary evaporators are now standard communal pieces of
equipment in the laboratory and the operating principle is that of a
reduced-pressure distillation except that the evaporation flask can be
rotated. This rotation reduces the risk of 'bumping', inherent in all
reduced-pressure distillations, and spreads the solution in a thin film on
the walls of the flask. This effectively increases the surface area of the
solution and increases the rate of evaporation, which is further enhanced
by the use of a vacuum.

Evaporation
~ When using a 'rovap' you must check that the re-

duced-pressure boiling point of the solute you are trying to isolate is be-
low the temperature of the water bath.
There are many variations in the details of the form of rotary film
evaporators and a typical assembly is illustrated in Fig. 17.1. A general guide
to the use of a rotary evaporator is given in Box 17.1.

Evaporation
When using rotary film evaporators you should take note of the following
safety advice:
• Never use flat-bottom flasks or conical flasks under reduced pressure (p.
110).
• Always check your flask for 'star' -cracks.
• Always make sure that your solution has cooled to room temperature
before you begin, otherwise it may boil vigorously and 'bump' when you
apply the vacuum, before it is lowered into the water bath.
• Do not rush to lower the flask into the water bath: wait to see what
happens to the extent of evaporation at room temperature.
• Always have the water bath just warm, not hot, at the start of the
procedure. If the water bath is too hot, allow it to cool or add cold water
or Ice.
(a)
• Check that all joints are 'sealed' and that the water pump is producing
a vacuum: it will change 'note' as the vacuum is produced, when it is
working properly. If there is no vacuum, the solution may not boil
and you will overheat it in trying to promote evaporation. The joints
may suddenly seal and the solution will then boil vigorously under the
reduced pressure and will 'bump' into the condenser and receiving
flask.
If it is necessary to evaporate volumes of solvent greater than the
capacity of the rotating flask, you can carry out the process batch-wise
involving several separate evaporations or the rotary evaporator can be
modified for continuous evaporation (Fig. 17.2). A thin Teflon® tube is
attached to the vacuum inlet adapter so that it feeds down the condenser
(b)
into the 'barrel' and another glass tube, dipping into the solution to be
Fig. 17.1 (a) Typical examples of a rotary evaporated, is connected to the air inlet on the vacuum adapter. Once the
evaporator; (b) exploded view of glassware. rotary evaporator is operating, the tap on the vacuum adapter is opened
a little. Solution is drawn up by the vacuum, runs into the rotating flask
and is evaporated. Careful control of the tap allows a constant volume of
solution to be sucked into the rotating flask and evaporated without
overfilling it.
Gas 'blow-down'
This procedure is useful for removing very small volumes of solvents
(about 2 mL) from solutes by blowing a stream of nitrogen over the
surface of the solution, while warming the solution gently. The main
application of the gas blow-down is in the isolation of small amounts of
solute from rotary evaporation or small-scale liquid-liquid extraction, for
further analysis by instrumental techniques, where the sample size may
be 20 mg or less: for example, infrared spectroscopy (p. 180), NMR
spectroscopy (p. 190), gas chromatography (p. 211) or liquid chromato-
graphy (p. 218).
The simplest system for evaporation by gas blow-down is shown in Fig.
17.3. A Pasteur pipette is connected by a flexible tube to a cylinder of
nitrogen, which has a gas blow-off safety system (p. 125). The sample is
placed in a special tube with a conical base, such as a ReactiVial®. Hold the
Pasteur pipette and direct a gentle stream of nitrogen towards the side of the
tube so that it flows over the surface of the liquid. As the solvent evaporates,
the liquid and tube will cool and may condense atmospheric water into the
tube. To prevent condensation, clamp the tube in a warm sand bath or above
a closed steam bath or in the hole of a purpose-designed aluminium heating

Evaporation
air or nitrogen
Pasteur pipette
\ r
Fig. 17.2 The procedure for continuous
solvent removal using a rotary evaporator.
Heating medium
Fig. 17.3 Gas 'blow-down' evaporation.
block, The following points should be noted when using a gas blow-down
system:
• Always carry out the operation in a fume cupboard.

• The solute should have negligible vapour pressure at room temperature,
otherwise it may eo-evaporate with the solvent.
• Do not heat the solution to boiling. Only apply enough heat to prevent
condensation of atmospheric water vapour.

------~0 Inert atmosphere methods
During your laboratory work you may need to carry out reactions using
chemicals which are described as air sensitive or moisture sensitive. These
compounds may react with water, oxygen, carbon dioxide and even nitrogen
(e.g. lithium metal). The most sensitive chemicals may require special
apparatus such as glove boxes or vacuum line equipment and you should
consult the appropriate specialist literature (Errington, 1997, p. 56). On the
other hand, many reactions involving some air-sensitive reagents (e.g.
organolithium compounds or hydride reducing agent) can be done on a small
scale using standard glassware with appropriate modifications. For simple
apparatus for inert atmosphere reactions, the basic requirements are:
• Inert atmosphere, usually nitrogen or argon, piped into the apparatus.

Nitrogen is the most commonly used inert gas, whereas argon is more
expensive but does have the advantage that it is denser than nitrogen and
is not lost from the apparatus as quickly. The inert atmosphere is
maintained in the apparatus by the use of a 'bubbler' (see p. 126) on one
of the outlets from the glassware - all other outlets must be stoppered or
capped with septa.
• Appropriate glassware for the experiment (see Chapter 16) which must be
dry.
low-pressure
gauge
• Dry solvents and chemicals (p. 127).
• Syringe techniques for dispensing and transferring chemicals to the
apparatus (p. 127).
high-pressure
gauge (cylinder)
Inert atmosphere
The source of the inert atmosphere is usually a cylinder of nitrogen or argon
diaphragm
regulator control
gas under pressure, which should be placed as close to the apparatus as
threaded bullet adapter possible to avoid long 'runs' of connecting rubber tubing.
to fit into cylinder %crease outlet
pressure
Gas cylinders
cylinder valve
The gas flow rate from the cylinder is controlled by the cylinder head
regulating valve (Fig. 18.1). Before you start make sure that the regulator
outlet tap is off (turn anti-clockwise until it feels 'free') and then open the
gas cylinder
valve to the cylinder with the cylinder spanner (turn anti-clockwise) and the
cylinder pressure should be indicated on the right-hand pressure dial. Switch
Fig. 18.1 The diaphragm pressure regulator.
on the gas at the regulator (turn slowly clockwise) until there is a reading on
the left-hand dial. Use the minimum pressure required to provide a steady
flow of gas. The gas flow rate from the regulator can be controlled further by
a needle valve on the regulator outlet, if one is fitted. To switch off, reverse
the instructions above.
Connection to the apparatus

Use clean, dry, thin-walled rubber tubing and special adapters with ground-
glass joints to connect the tubing to your reaction flask or to the inlet pipe of
a 'bubbler'. Where a single cylinder supplies several outlets, e.g. in a fume
cupboard, the gas flow rate may change markedly when someone turns off
one of the outlets, resulting in an increase in gas pressure to your equipment.
You should, therefore, fit a gas 'blow-off valve between your gas supply and

Inert atmosphere methods
the apparatus. To do this, fit a glass tube 'T-piece' in the gas line to your
apparatus and connect it to a Dreschel bottle containing mineral oil to a
depth a little more than that of the mineral oil in the 'bubbler'. If there is a
sudden upward change in gas pressure, the gas will be vented through the
Dreschel bottle, as well as through your apparatus.
Gas 'bubblers'
The exit of the inert gas from the apparatus must be protected by a gas
'bubbler'. The 'bubbler' allows you to monitor the flow of inert gas through
the apparatus and prevents the entry of air into the apparatus. Several
designs of gas 'bubbler' are available (Fig. 18.2) and it is usual to connect the
'bubbler' to the apparatus at the top of the condenser. You should make sure
that the 'bubbler' contains enough mineral oil to create a seal from the
atmosphere and that it has a bulb above the mineral oil to collect any
mineral oil, which could be sucked back into your apparatus if there is a
sudden contraction in the volume of inert gas in the apparatus. Such changes
in volume will occur if you suddenly cool the apparatus without increasing
the gas flow to compensate for the resulting reduction in inert atmosphere
volume.
Apparatus
Depending upon the type of reaction to be carried out, one of the assemblies
shown on p. 119 may be used, with additional modifications for inert gas
inlet and outlet. You should consider very carefully what is required: heating
or cooling, magnetic or mechanical stirring, temperature measurement, etc.
The gas inlet can be either directly into the reaction flask or into the inlet
arm of the gas 'bubbler' - it depends on the number of 'necks' available on
the reaction flask.
seal or
inert -.-
gas in
Drying glassware
All equipment to be used should be dried (e.g. in an oven overnight at 125 QC-
do not forget to remove all plastic or TeflonC1\lcomponents before placing the
glassware in the oven). After drying, the apparatus should be greased,
-.- inert
gas out
assembled hot, using heat-resistant gloves as protection, and allowed to cool
with the inert gas flowing rapidly through it. Once cool, the water connections
for the condenser should be fitted - screw-on water connectors (p. 118) are
most useful in this context.
inert gas out
t inert-.-
gas In Addition of chemicals
Chemicals should be added to the reaction flask using a pressure-
-.- inert
gas out
equalizing dropping funnel (p. 119). Liquids and solid compounds are best
added as solutions in the solvent used in the reaction. If the solid is
insoluble, a little solvent should be added to the reaction flask, the
'bubbler' outlet sealed, a stopper to the flask opened and the gas flow rate
increased. The solid can then be added from a wide-stemmed filter funnel,
protected by the inert gas and solvent vapour, so that air does not enter
the apparatus. Then, simultaneously unseal the 'bubbler' and restopper the
t
inert gas in
flask, and then turn down the gas flow. Alternatively a solids addition tube
(Errington, 1997, p. 124; Harwood, et al., 2000, p. 89; Furniss, et al., 1989,
Fig. 18.2 Typical 'bubblers'. p. 82) can be used.

Solvents and chemicals

All chemicals and solvents used in inert atmosphere reactions must be dry.
Most of these materials provided by suppliers are not dry enough, even
solvents which you consider to be immiscible with water, and may contain
enough moisture to hinder the reaction or reduce the yield of your product.
Therefore you must ensure that all chemicals to be used in the process have
been dried to the appropriate levels, as described below.
Solid chemicals
These should be dried by the methods outlined on p. 39. The most common
approach is to dry the chemical in an oven and then allow it to cool in a
vacuum desiccator (p. 40). Techniques for extremely air-sensitive solids can
be found in the specialist literature.
Liquid chemicals
All liquids should be dried by a method appropriate to the amount of
water they may contain (p. 41). Generally, the liquid should be dried with
a solid drying agent (p. 41) which does not react with the chemical
(consult the appropriate literature or your instructor), filtered, distilled
(p. 107), then stored over molecular sieves (p. 41) in a bottle capped by
a septum and redistilled before use. Alternatively, the liquid can be dissolved
in a solvent, the solution dried (p. 41), the solvent removed by evaporation
(p. 121) and the liquid distilled and stored as described above.
Solvents
The solvent will have the greatest volume in your reaction and it must be
dry. Most laboratory-grade solvents, as supplied by manufacturers, contain
varying amounts of water and therefore must be dried by the appropriate
method before use. If you are required to dry the solvent, you should
consult the literature (Errington, 1997; Harwood, et al., 2000; Furniss, et aI.,
1989).
Some manufacturers supply dry solvents in 2.5 L quantities for inert
atmosphere reactions and HPLC (p. 2 I8). These solvents are relatively
expensive but may be economic in terms of time and expense if one-off
reactions are required. However, such solvents should be treated with
suspicion if the containers are less than half-full, since air and moisture may
have been allowed into the container by previous users. If you have any
doubts, dry the solvent.
Syringe techniques
Many air-sensitive chemicals are supplied as solutions in nitrogen-filled
bottles, which are sealed by a septum, and small volumes (up to 25 mL) of
these solutions are best transferred to the apparatus using glass syringes.
Similarly, air-sensitive liquids can be added to the reaction using a syringe.
~ When removing air-sensitive reagents from nitrogen-

filled bottles, you must replace the volume of liquid removed with inert
gas (nitrogen) from a gas cylinder or balloon, via a needle, otherwise air
(water, oxygen and carbon dioxide) will be pulled into the bottle as a re-
sult of the vacuum you have created.

Syringes
Glass, gas-tight syringes with a Luer lock fitting are the most versatile type of
syringe and they come in a range of sizes. The Luer lock enables the stainless
steel needle to be locked in place on the end of the syringe so that there is no
danger of the needle dropping off the syringe during the transfer process
(Fig. 18.3). Variations in syringe types include those with Teflon® -tipped
pistons (plungers), which are somewhat more expensive.
Before using a syringe, always check that it is working by sucking up a little
~lllllllllJilllllllll of the solvent to be used, ensuring that air is not sucked into the syringe either
via the Luer lock or down a gap between the syringe and piston. If all is correct,
glass barrel glass piston
disassemble the syringe and needle, dry in an oven at 120 QC (not if Teflon®
tipped) and allow to cool in a desiccator. Once you have transferred the air-
~lllil'lill'iIlJilill sensitive reagent, you must clean out the syringe and needle immediately, by the
appropriate method, as air will get into the needle and syringe and decompose
glass barrel
the reagent causing the syringe to jam or the needle to block.
Teflon® seal Needle-to-tubing connectors

------++ntnallmlnllhmJmllmmmlmdnnl ~~~
These adapters allow a Luer lock syringe needle to be connected to rubber
needle j =========='o tubing (Fig. 18.4). An inert gas supply can then be piped, via the needle, into
locking nut " ® stainless steel piston a bottle to allow the transfer of large volumes of solvent or air-sensitive
Tellon R tip
reagents to the apparatus. Alternatively a balloon can be taped to a short
Fig. 18.3 Syringes for inert atmosphere work. piece of thick-walled rubber pressure tubing to provide a supply of nitrogen
when withdrawing air-sensitive reagents using a syringe (Fig. 18.5).
balloon
tape
thick-walled rubber tubing
needle-ta-tubing connector
---- Luer-Iocklit® needle

Fig. 18.4 Needle-to-tubing connector.
(a)
-
ring support to hold
bottle securely
reagent bottle
(b)
Fig. 18.5 Inert atmosphere transfers: (a) balloon and needle; (b) preserving the
inert atmosphere while removing reagent.

Syringe needles and cannulae

Stainless steel Luer lock syringe needles come in various lengths and
diameters. The length of needle you will need depends on the size of the vessel
from which you wish to withdraw the liquid; the diameter required depends
on the size of the syringe - you should not use a large-diameter needle with a
small-volume syringe - and the viscosity of the solution or liquid. Needle
diameters are expressed in 'gauge': the higher the gauge, the narrower the
needle diameter. For most inert atmosphere work you should use a needle
with a 'non-coring' or 'deflecting' tip (Fig. 18.6), which ensures that a piece of
the septum is not trapped in the needle when you push it through. Cannulae
are long, flexible, double-ended needles made from stainless steel or inert
plastics, which are used to transfer large volumes of reagents or solvents from
one vessel to another under inert gas pressure (Fig. 18.7).
A generic method for transferring an air-sensitive reagent, by syringe, from
a bottle to a pressure-equalizing dropping funnel is described in Box 18.1.
inert gas
)<"
Fig. 18.6 Needle with non-coring tip for

piercing septa, ~ ring clamp
Fig. 18.7 Transfer of air-sensitive reagents using the double-ended needle technique.
seal with septum

-
septum
pressure-equalizing
clamp addition funnel
condenser
clamp
three-neck flask
'flea'
,\1//
__ ....
-@- stirrer
Fig. 18.8 Transferring an air-sensitive reagent to a pressure-equalizing dropping

funnel.


Resources
Resources for laboratory techniques
Books
Bennett, S.W. and O'Neale, K. (1999) Progressive Development of Practical
Skills in Chemistry. A guide to early-undergraduate experimental work, Royal
Errington, R.J. (1997) Advanced Practical Inorganic and Metalorganic
Chemistry, Blackie Academic and Professional, London.
Furniss, BA., Hannaford, A.J., Smith, P.W.G. and Tatchell, A.R. (1989)
Vogel's Textbook of Practical Organic Chemistry, 5th Edn, Longman,
Harlow, Essex.
Halpern, A.M. (1997) Experimental Physical Chemistry, Prentice Hall,
Harlow, Essex.
Harwood, L.M., Moody, C.l and Percy, J.M. (2000) Experimental Organic
Chemistry, 2nd Edn, Blackwell Science Ltd, Oxford.
Lehman, J.W. (1999) Operational Organic Chemistry. A problem-solving
approach to the laboratory course, 3rd Edn, Prentice Hall, Harlow, Essex.
Lister, T. (1996) Classic Chemistry Demonstrations, Royal Society of
Loewenthal, H.J.E. (1990) A Guide for the Perplexed Organic Experimentalist,
2nd Edn, John Wiley and Sons Ltd, Chichester.
Mendharn, J., Denney, R.e., Barnes, J.D. and Thomas, M.J.K. (2000)
Harlow, Essex.
Nelson, J.H. (1997) Laboratory Experiments for Chemistry, Prentice Hall,
Harlow, Essex.
Sharp, J.T., Gosney, I. and Rowley, A.G. (1989) Practical Organic
Chemistry, Chapman and Hall, London.
Suib, S.L. and Tanaka, J. (1999) Experimental Methods in Inorganic
Chemistry, Prentice Hall, Harlow, Essex.
Williamson, K.L. (1999) Macroscale and Microscale Organic Experiments,
3rd Edn, D.C. Heath and Company, Lexington.
Videos
Basic Laboratory Skills, LGC, Royal Society of Chemistry, Cambridge
(1998).
Further Laboratory Skills, LGC, Royal Society of Chemistry, Cambridge
(1998).

-8 Qualitative techniques for inorganic
analysis
Qualitative techniques are used to identify cations and anions in aqueous

solution by simple reactions, mostly involving the production of a precipitate,
evolution of gas or a visual colour change. It is important to make
observations accurately and to interpret them in a step-wise manner.
The following basic equipment is required to carry out qualitative analysis:
• Test tubes - in which the reactions are performed.

• Cork or rubber stoppers - for the protection of the contents of test tubes
from contamination and for safe storage.
• Test tube rack - this allows test tubes to be stored upright when not in
use.
• Test tube holder - this allows individual test tubes to be heated safely.
• A glass rod - this has several functions including the stirring, transfer of
solutions, and the break-up of precipitates.
• Watch-glasses these have several functions including the covering of
beakers to prevent contamination and as a receptacle for solutions.
• A wash bottle containing distilled water.
• Spatula - for transferring small quantities of solids.
• Pasteur pipettes - for transferring liquids.
• Micro-Bunsen burner - for heating solutions to boiling and for
evaporating solutions.
• Evaporation crucible - this is used as a container for solutions when
complete evaporation of liquid is required, leaving a solid product.
• Crucible tongs - for removing the crucible from the heat source.
• Centrifuge - for separating precipitates from solution.
• Heated water bath.
Table 19.1 Typical reagents for qualitative
analysis
Reagents
At the start of your experimental work always check that the appropriate
reagents are readily available (a list of commonly used reagents is given in
Table 19.1). Note that it is essential to use distilled water in all qualitative
analysis. Tap water contains ions such as Ca2 +, Mg2 +, Fe2 +, Fe3 +, SO~-
and Cl- and its use could lead to 'false-positive' results.
Testing for anions and cations

Specific literature containing tests for the determination of anions and cations
can be found in the resources section (p. 160). In general, however, the
following tips are useful when carrying out qualitative analysis:
• Always work tidily to prevent cross-contamination of samples.

• Ensure that all glassware has been cleaned thoroughly in detergent and
then rinsed twice with distilled water. Invert the test tubes to drain; never
dry the inner surface with towelling or tissue.
• Label test tubes at the start - it may prove difficult to remember what
you have done later on.
Classical techniques 135

Qualitative techniques for inorganic analysis
Qualitative tests for cations and anions • Always test solutions with a known composition before you attempt to
An unknown solution was tested as follows: analyse solutions with an unknown content. This allows you to gain the
necessary experience in solution manipulation, observation skills and the
Test Observation Conclusion
interpretation of results.
2 drops of dilute White Sulphate
HCI, boil, then add 1 precipitate ISO~-I present
• The colour of solutions and/or precipitates has to be interpreted from
drop BaClz solution written or oral information. Interpretation of colour can be subjective, so
you will need to gain sufficient experience using solutions of known
content to establish how a particular colour appears to you.
• Establish a protocol for recording of observations after carrying out
different tests (Fig. 19.1).
Test performed on
unknown solution • Reagents should be added from Pasteur pipettes held with the tip just
above the mouth of the test tube. Never put Pasteur pipettes inside test
Fig. 19.1 Recording your observations in tubes as this can lead to contamination of the reagents.
qualitative analysis. • Effective mixing of the test solution and added reagents is essential. This
can be achieved by holding the test tube between the thumb and index
finger of one hand and 'flicking' the tube with the index finger of your
other hand. Alternatively, solutions can be mixed by bubbling air from a
Pasteur pipette held at the bottom of the test tube.
• Evolved gas can be drawn up into a Pasteur pipette and then bubbled
through a test solution, e.g. CO2 can be drawn into a Pasteur pipette and
then 'blown' out through lime water (Ca(OH)2 solution) giving a milky-
white solution.
• Solutions can be tested for pH using litmus paper. Never place litmus
paper directly into the test solution. Instead, dip a glass rod into the
solution, remove, touch the wet glass rod onto the litmus paper and note
the colour. Acidic solutions change blue litmus paper to red; alkaline
solutions change red litmus paper to blue. Alternatively, universal
indicator paper can be used. In this case, the orange paper turns 'reddish'
with acidic solutions and 'bluish' with alkaline solutions. By comparing
any change in colour with a chart (supplied with the universal indicator
paper), the pH of the solution can be estimated.
Centrifugation of solutions
Centrifugation causes particulate material to accumulate at the bottom of the
test tube. The procedure for centrifuging your sample is described in Box
19.1. The speed and time of the run will depend on the centrifuge available,
but will typically be in the range 5000-10 000 rpm for 5-10 min, respectively.
Always allow the centrifuge to stop in its own time, as abruptly halting the
centrifuge will disturb light precipitates. After centrifugation, hold the test
tube at an angle so that it is easy to remove the liquid component (or
centrifugate) with a Pasteur pipette (Fig. 19.2). You will find that it is
difficult to remove all the centrifugate from the precipitate, and to maximize
the transfer of centrifugate it is necessary to wash the precipitate. This is
carried out as follows:
residue • Add a small quantity of distilled water to the precipitate.

• Use a glass rod to break up the precipitate and mix thoroughly.
• Recentrifuge the mixture.
• Transfer the liquid obtained to the original centrifugate and store this
liquid solution for further analysis.
Fig. 19.2 Separation of liquid from a residue • Repeat the washing process, but this time discard the centrifugate, and
using a pipette. retain the precipitate for further tests.
136 Classical techniques

Heating test tubes and other containers

It is often necessary to heat a solution in a test tube, either to cause
precipitation or to dissolve a precipitate. You can carry out this heating
effectively and safely by partially immersing the test tube containing the
mixture in a simmering boiling-water bath (remember to use a test tube
holder!).
It is possible to reduce the volume of the solution in the test tube, i.e. to
pre-concentrate the sample, by evaporation. Two different methods can be
employed.
1. Transfer the solution to a small evaporating dish. Place the evaporating

dish on a wire gauze located on a tripod stand, and apply heat using a
micro-Bunsen burner. Note that the volumes of solutions in qualitative
analysis are often small, and excessive heating might result in hardening
of any residue, making it unusable.
2. Alternatively, evaporate the solution directly in a test tube by gentle
heating over a micro-Bunsen burner. Remember to use a test tube
holder. Position the test tube at an angle with the tip of the Bunsen
burner flame positioned at the upper surface of the liquid. Place a
glass rod inside the test tube and rotate constantly. This acts to
disperse bubbles of steam that are given off. Extreme caution is
required with this method of evaporation, as the steam bubbles can
cause the solution to 'bump' out of the test tube. 'Bumping' can
result in hot (and maybe toxic) substances being ejected over a
suprisingly large distance. To prevent this it is normal to add anti-
bumping granules (p. 31).

Table 19.2 Flame tests for cations
Fig. 19.3 Holding a nichrome wire in a flame test.
Flame tests
Simple flame tests can be carried out on solid samples. Place a little of the
* The colour is often obliterated by trace
impurites of sodium present (sodium gives
solid on a watch-glass and moisten with a drop of concentrated hydrochloric
an intense yellow colour). You can overcome acid. The purpose of the hydrochloric acid is to produce metal chlorides
this by viewing the colour through cobalt-blue which are volatile at the temperature of the Bunsen burner.
glass which allows the lilac colouration from Pre-clean a platinum or nichrome wire by holding it in the hottest part of
potassium to be seen.
the Bunsen flame (just above the central blue cone) until there is no coloured
** Viewing through cobalt-blue glass also
flame from the wire. Cool, then dip the cleaned wire into the moistened solid
allows calcium and strontium to be
distinguished. In this case, calcium is light sample. Place the wire at the edge of the Bunsen flame (Fig. 19.3) and record
green in colour while strontium appears purple. the colour of the flame from the sample (see Table 19.2).

-------0 Gravimetry
Gravimetric analysis is the process of converting an element into a definitive

compound, isolating this compound from other constituents in a sample and
then weighing the compound (Box 20.1). The weight of the element can then
be calculated from the formula of the compound and the relative atomic
masses of the elements involved. You need to be able to weigh accurately, by
difference, a substance to four decimal places (see p. 23).
~ The essential component of gravimetric analysis is

the transformation of the element of interest into a pure stable solid
compound, suitable for weighing.
The most common approach for isolating the element is by precipitation

from a solution where it is present in ionic form (see p. 50). Ideally, the
constituent under investigation is precipitated out of solution as a water-
insoluble compound, so that no losses occur when the precipitate is separated
by filtration, washed free of soluble impurities and then weighed.

Gravimetry
Precipitation
Table 20.1 Common precipitants Inorganic ions can be separated from mixtures using organic reagents
(precipitants), with which they form sparingly soluble, often coloured,
compounds. The precipitants usually have high molecular weights, so a small
quantity of the ion will produce a large amount of precipitate. Ideally, the
Dimethylglyoxime Ni2+ Pd 2+, Pt2+, Bi3+ precipitant should be specific for a particular ion, though this is rarely so.
and Au3+ Examples of common precipitants and their target ions are shown in Table
Cupferron Sn4+ Cu2+ and Pb2+ 20.1 and their structures are shown in Fig. 20.1.
3
8-Hydroxyquinoline A1 + Many metals Dimethylglyoxime is only slightly soluble in water (0.40 g L -I) and it is
(oxinel
therefore used as a 1% wjv solution in ethanol. Cupferron, the ammonium
salt of N-nitroso-N-phenylhydroxylamine, is used as a 5-10% wjv solution in
hydrochloric acid or sulphuric acid. Oxine (8-hydroxyquinoline) is almost
CH3-y~NOH
insoluble in water and is used as either a 2% or a 5% wjv solution in
CH3-C~NOH
2 mol L -1 acetic acid.
Dimethyloglyoxime
When precipitating a compound:
...-NO
(QrN"O-NHt • Mix your reagents slowly, with continuous stirring, to encourage the
growth of large crystals of the compound.
Cupferron
• Improve the precipitation process by heating your solutions: ideally, one
~ or both solutions should be heated to just below boiling point.
yN:/
• Wash your precipitate with a dilute electrolyte solution, to remove any
OH
other constituents (it is essential to remove any impurities). Choose a
8-Hydroxyquinoline
solution that does not interact with the precipitate, and that is volatile at
Fig. 20.1 Structures of common precipitants. the drying temperature to be used.
• Use the minimum quantity of wash solution as no precipitate is absolutely
-- Gooch crucible insoluble. While suitable wash solutions include dilute electrolytes, e.g.
rubber cone
ammonium salts, ammonia solution or acids, pure water is rarely used, as it
Gooch funnel
may dissolve the precipitate.
• Test your filtered wash solution for impurities using simple qualitative
tests (Chapter 19). Continue until your final washing solution contains no
Buchner flask trace of other constituents.
• It is best to wash repeatedly with several small amounts of solution,
allowing the precipitate to drain between washings.
Filtration
Fig.20.2 Experimental arrangement for To carry out this procedure, you will need to assemble a Gooch crucible and
filtration of precipitate. funnel on a Buchner flask, clamped for stability using a retort stand (Fig. 20.2).
The glass Gooch crucible has a porous disk of sintered ground glass, typically
of pore diameter 20-30 !lm, which is satisfactory for moderately sized
precipitates. Fit the crucible into a Gooch funnel using a rubber cone, put the
funnel into a one-holed rubber bung and then into a Buchner flask. The tip of
the funnel must project below the side arm of the flask to prevent loss of filtrate
down the side arm (see p. 28). Then, connect the Buchner flask to a water
pump. Pour your precipitate suspension into the Gooch crucible, using a glass
rod (p. 18) to direct the liquid into the centre of the sintered base. The lower
end of the glass rod should be close to, but not touching, the sintered-glass
base. Never overfill the crucible. The precipitate remaining in the bottom of the
beaker should be rinsed out with the filtrate solution: disconnect the pump and
pour the filtrate back into the beaker containing the precipitate. The pump
should be disconnected by pulling off the vacuum tube from the Buchner flask.
Fig. 20.3 Structure of nickel-dimethylglyoxime On no account turn off the water pump while doing this. You may need to
complex. rinse the beaker several times, to collect all of the precipitate in the crucible.

~------0
Procedures in volumetric analysis
Volumetric analysis, also known as titrimetric analysis, is a quantitative

technique used to determine the amount of a particular substance in a
solution of unknown composition.
This requires:
• A standard solution, which is a solution of a compound of accurately

known concentration, that reacts with the substance to be analysed.
• The test solution, containing an unknown concentration of the substance
to be analysed.
• Some means of detecting the end-point of the reaction between the
standard and test solutions, e.g. a chemical indicator or, in the case of
potentiometric titrations, a pH electrode (see Chapter 34). Some reactions
exhibit a colour change at the end-point without the addition of an
indicator.
The volume of standard solution that reacts with the substance in the test
solution is accurately measured. This volume, together with a knowledge of
concentration of the standard solution and the stoichiometric relationship
between the reactants, is used to calculate the amount of substance present in
the test solution. Specific examples of the different types of calculations
involved are shown in Chapters 22 to 25.
Classification of reactions in volumetric analysis

There are four main types of reaction
1. Acid-base or neutralization reactions, where free bases are reacted with a
standard acid (or vice versa). These reactions involve the combination of
hydrogen and hydroxide ions to form water.
2. Complex formation reactions, in which the reactants are combined to
form a soluble ion or compound. The most important reagent for
formation of such complexes is ethylenediamine tetra-acetic acid, EDTA
(as the disodium salt).
3. Precipitation reactions, involving the combination of reactants to form a
precipitate.
4. Oxidation-reduction reactions, i.e. reactions involving a gain (reduction)
or loss (oxidation) of electrons. The standard solutions used here are
either oxidizing agents (e.g. potassium permanganate) or reducing agents
(e.g. iron (11)compounds).
What can be measured by titration?

• The concentration of an unknown substance e.g. 0.900 mol L -I.
• Percentage purity, e.g. 56%.
• Water of crystallization, e.g. (NH4hS04.nH20.
• Percentage of a metal in a salt, e.g. 12% Fe in a salt.
• Water hardness, e.g. determination of the concentration of calcium and
magnesium ions.
Examples of the types of calculations used in volumetric analysis are shown
in Box 21.1.


Titrations
The process of adding the standard solution to the test solution is called a
titration, and is carried out using a burette (see below). The point at which
the reaction between the standard solution and the test substance is just
complete is called the equivalence point or the theoretical (or stoichiometric)
end-point. This is normally detected by a visible change, either of the
standard solution itself or, more commonly, by the addition of an indicator.
Standard solutions
A standard solution can be prepared by weighing out the appropriate
amount of a pure reagent and making up the solution to a particular volume,
as described on p. 24. The concentration of a standard solution is expressed
in terms of molarity (p. 19). A substance used in a primary standard should
fulfil the following criteria:
• It should be obtainable in high purity (> 99.9%).
• It should remain unaltered in air during weighing (i.e. it should not be
hygroscopic).
• It must not decompose when dried by heating or vacuum.
• It should be capable of being tested for impurities.
• It should be readily soluble in an appropriate solvent.
• It must react with the test substance stoichiometrically and rapidly.
Preparing a standard solution

The molarity of a solution is the concentration of the solution expressed as
mol L:": If xg of a substance of molecular weight M, is dissolved in ymL of
distilled water, the moles of substance dissolved = ; = m. Therefore,
. m x 1000 r
molanty (mol L:") =--- [21.1]
Y

Primary standards prepared from solid compounds should be weighed out

using the 'weighing by difference' method as described in Chapter 4, and
accurately made up to volume using a volumetric flask. Complete transfer of
the substance from the weighing vessel to the volumetric flask is best
achieved by inserting a funnel into the neck of the flask (Fig. 21.1). As much
of the solid as possible should be transferred via the funnel. The funnel
should be washed with distilled water prior to removal. Distilled water is then
added to the flask, with occasional swirling to help to dissolve the solid. This
is continued until the meniscus is about 1 cm below the volume mark. At this
point a stopper is inserted and the flask is inverted several times to ensure the
solid is completely dissolved. Finally, using a Pasteur pipette, distilled water
is added up to the volume mark. The solution should be thoroughly mixed
before use.
If the solid is not readily soluble in cold water it may be possible to
dissolve it by stirring in warm water in a beaker. After allowing the solution
to cool to room temperature, it can be transferred to the volumetric flask
using a glass rod and filter funnel (Fig. 21.1) followed by several rinses of the
glass rod/filter funnel. Finally, the solution is made up to the mark with
distilled water.
Filling a burette
• Clamp a clean 50.00 mL burette (p. 10) in a laboratory stand. Place a
beaker on a white tile immediately below the outlet of the burette (Fig.
21.2).
• Place a small filter funnel on top of the burette and, with the tap open,
carefully pour in the standard solution (or titrant) until it starts to drain
into the beaker.
• Close the burette tap, and fill the burette with the standard solution until
the meniscus is about 1-2 cm above the zero mark. Remove the funnel.
• Open the tap and allow the solution to drain until the meniscus falls to
the zero mark. The burette is then ready for the titration.
Note that to avoid contamination the solution in the beaker should be
discarded, rather than recycled.
Using a pipette
A clean 2S.00mL pipette (p. 10) is required together with a suitable pipette
filler. Various designs of pipette filler are available. The most common type is
based on a rubber-bulb suction device. It is best to evaluate a range of
pipette fillers, if available in the laboratory, for ease of use and performance.
The pipetting procedure is as follows:
• Pour the solution of unknown composition (the titrand) into a beaker.
Never place the pipette in the volumetric flask containing the solution as
this can lead to contamination of the solution from the external surface
of the pipette.
• Using your pipette filler, draw the titrand to just beyond the graduation
mark (Fig. 21.3a). Remove the pipette filler, and invert the pipette to
allow the solution to drain out. This ensures that the titrand used
subsequently will be undiluted and uncontaminated by any residue or
liquids in the pipette.

----- burette
6
6 clamp
8
retort stand
rinse~
solution
Fig. 21.1 Quantitative transfer of a solid to

a volumetric flask. Fig. 21.2 Apparatus for a titration.
• Refill the pipette until the meniscus of the titrand is above the graduation
mark (Fig. 21.3a). Remove the pipette filler and block the hole at the top of
the pipette with the index finger of your right hand (if right-handed; Fig.
21.3b). Carefully raise the pipette to eye level, and allow the titrand to drain
out into a beaker by lifting your finger slightly from the top of the pipette.
Continue until the bottom of the meniscus is on the graduation mark.

• Wipe the outside of the pipette with a tissue (Fig. 2l.3c). Be careful not
to touch the point of the pipette with the tissue otherwise solution will be
lost by capillary action.
• Allow the pipette's contents to drain into a conical flask.
• Finally, touch the end of the pipette on the wall of the flask (Fig. 21.3d),
and rinse the inside of the neck of the flask with distilled water. This will
ensure that exactly 25.00 mL of the test solution has been delivered by the
pipette.
Note that it is normal for a small quantity of solution to remain in the

pipette tip. This volume is taken into account when pipettes are calibrated, so
do not attempt to 'blowout' this liquid into the conical flask.
Performing a titration
Add one or two drops of indicator to the titrand contained in the conical
flask. For a right-handed person the process is as follows:
(a) (bl
• Hold the conical flask containing the tit rand and the indicator in your
right hand, and control the tap of the burette with your left hand. The
burette should be arranged so that the tap is on the opposite side of the
burette to your palm. In this way, your left hand also supports the body
of the burette (Fig. 21.4).
• Add the titrant by opening the tap, and simultaneously swirl the contents
of the conical flask. This may take a bit of practice, so do not worry if
you cannot do it straight away. The titrant can be added quickly at first,
but as the end-point approaches, additions should be made drop-wise.
• The end-point is indicated when the appropriate colour change takes
place. When the end-point is reached, one drop of titrant should be
sufficient to cause the colour change. You should note the volume used
for the titration to the nearest 0.1-D.05mL. This is done by reading off
the volume of titrant used (Fig. 21.5).
• Refill the burette to the zero mark using a funnel ready for the next
titration.
(c) (d)
Fig. 21.3 Using a pipette.

Volumetric analysis - calculating the concentration
of a test substance
The calculation should be carried out in a logical order as follows:
1. Write the balanced equation for the reaction between the standard and
the test substance.
2. From the stoichiometry of the reaction, determine how many moles of
the test substance react with 1 mole of the standard substance. For
example, in the reaction between an H2S04 standard solution and an
NaOH test solution:
[21.2]
Therefore 2 moles of NaOH react with 1 mole of H2S04.

3. Calculate the number of moles of standard substance used to reach the
end-point of the reaction. This can be determined from knowledge of the
concentration of the standard solution (mol L -1) and the volume of
titrant used (mL). Remember to take care with units - in this instance
division by a factor of 1000 is required to convert mL to L:

Fig. 21.5 Reading a burette. Place a white card

below the level of the meniscus. This allows an
accurate reading to be made.
Fig.21.4 Performing a titration.
concentration (mol L -1) x volume (mL)

N urn b er 0 f mo Ies = 1000
4. The number of moles of test substance present in the titrand is then

obtained from knowledge of the equivalences. In the example given
above (point 2) the number of moles of test substance is twice the
number of moles of standard substance. Therefore, if X moles of H2S04
are used (as calculated in point 3), 2X moles of NaOH were present in
the initial volume of test solution.
5. Finally, the concentration of the test solution can be calculated using the
formula:
Concentration of test lOOOxamount of test substance (mol)
solution (mol L -I) = initial volume of test solution (mL)
Again, the factor of 1000 is used to convert mL to L.

------~0 Acid-base titrations
The titration of an acid solution with a standard solution of alkali will

determine the amount of alkali which is equivalent to the amount of acid
present (or vice versa). The point at which this occurs is called the
equivalence point or end-point. For example, the titration of hydrochloric
acid with sodium hydroxide can be expressed as follows:
NaOH(aq) + HCl(aq) --+ NaCl(aq) + HzO(I) [22.1]
If both the acid and alkali are strong electrolytes, the resultant solution will be
neutral (pH 7). If on the other hand either the acid or alkali is a weak
electrolyte the resultant solution will be slightly alkaline or acidic, respectively.
In either case, detection of the end-point requires accurate measurement of
pH. This can be achieved either by using an indicator dye, or by measuring the
pH with a glass electrode (described in Chapter 7).
Acid-base indicators
thymol blue Typical acid-base indicators are organic dyes that change colour at or near
the equivalence or end-point. They have the following characteristics:
• They show pH-dependent colour changes.

• The colour change occurs within a fairly narrow pH range (approximately
2pH units).
• The pH at which a colour change occurs varies from one indicator to
another, and it is possible to select an indicator which exhibits a distinct
red yellow colour change at a pH close to the equivalence or end-point.
«pH 1.7) (pH 1.7-89)
Selected common indicators together with their pH ranges and colour

phenolphthalein changes are shown in Table 22.1. Examples for thymol blue and
phenolphthalein are shown in Fig. 22.1.
Table 22.1 Colour changes and pH range of selected indicators
colourless red Thymol blue 1.2-6.8 Red Yellow

«pH 61 (pH 8.3-10)
Methyl orange 2.8-4.0 Red Yellow
Figure 22.1 Examples of indicators used in
Methyl red 4.3-6.1 Red Yellow
Phenol red 6.8-8.2 Yellow Red
acid-base titrations: thymol blue and Phenolphthalein 8.3-10.0 Colourless Pink/red
phenolphthalein
15
Neutralization curves
10 A plot of pH against the volume of alkali added (mL) is known as a
:c
0.
neutralization or titration curve (Fig. 22.2). The curve is generated by a
'potentiometric titration' in which pH is measured after each addition of
alkali (or acid). The significant feature of the curve is the very sharp and
sudden change in pH near to the equivalence point of the titration. For a
50 100 150 ZOO strong acid and alkali this will occur at pH 7. If either the acid or base
volume of NaOH added (mLI concentration is unknown, a preliminary titration is necessary to find the
Figure 22.2 A typical neutralization curve: 0.1 approximate equivalence point followed by a more accurate titration as
M HCIwith 0.1 M NaOH. described on p. 146. The ideal pH range for an indicator is 4.5-9.5.

Acid-base titrations
14 Determination of the equivalence point

From the neutralization curve (Fig. 22.2), the initial and final slopes are
12 drawn (Fig. 22.3) and a parallel line is drawn such that the mid-point is
on the curve. This is the equivalence point, producing a titration value of
10
xmL.
:I:
c.
Example calculations
Standardization of a sodium hydroxide solution
What is the molarity of a solution of sodium hydroxide, 25.0 mL of which
2 requires 21.0 mL of a standard solution of hydrochloric acid of concentration
0.100 mol L -I for neutralization?
50 100 150 200
Following the sequence in Box 21.1:
volume of NaOH added ImL) 1. Write the equation:
Figure 22.3 Determination of the equivalence NaOH(aq) + HCI(aq) -+ NaCI(aq) + H20(l) (22.2)
point.
2. Determine the equivalences.
Equation [22.2] shows that I mole of NaOH requires 1 mole of HCI for
neutralization, i.e. 1 mole of NaOH is equivalent to I mole of HCI.
3. Calculate the number of moles of standard used.
Number of moles concentration (mol L -I) x volume (mL)
of HCl 1000
0.100 molL -1 x 21.0mL
1000
= 2.1 X 10-3 mol
4. Calculate the number of moles of NaOH in the initial volume of test

solution.
As indicated in point 2 above, 1 mole of NaOH is equivalent to 1
mole of HCI. Therefore:
no. of moles of NaOH in initial volume = no. of moles of HCl used

= 2.1 X 10-3 mol
5. Determine the concentration of the NaOH solution.

Concentration of test 1000 x amount of test substance
solution (mol L -1) initial volume of test solution (mL)
1000 x 2.1 x 1O-3(mol)

25 (mL)
= 0.084molL-1
Standardization of a sodium hydroxide solution using potassium

hydrogen phthalate as a primary standard (p. 143)
An accurately weighed amount (5.1100 g) of potassium hydrogen phthalate
(KHCgH404) was dissolved in distilled water (250.00 mL). This solution
(25.00mL) required sodium hydroxide solution (23.50mL) to reach
equivalence. What is the molarity of the sodium hydroxide solution? (Note
that, in this case, 25.00mL of the standard solution was used as the titrand,
whereas the test solution (NaOH) was the titrant.)

Acid-base titrations
Following the sequence in Box 21.1:

1. Write the equation.
NaOH(aq) + KHCgH404(aq) --> NaKCgH404(aq) + H20(l) [22.3]

2. Determine the equivalences.
Equation [22.3] shows that 1 mole of NaOH is equivalent to 1 mole of
KHCgH404·
3. Calculate the number of moles of standard used.
Firstly, the concentration of the standard solution of potassium
hydrogen phthalate must be calculated.
The molecular weight of KHCgH404 is 204.22gmol-l. Therefore
5.1100g of KHCgH404 is equivalent to 5.1100 (g)/204.22 (g mol ") =
0.025 mol.
We can now calculate the concentration of the standard solution:
Concentration of standard 1000 x amount of KHCgH404 (mol)

solution (mol L -I) volume of standard solution (mL)
1000 x 0.025 (mol)
250(mL)
= 0.100moIL-1
The number of moles of standard used in the titration is as follows.
Number of moles concentration (mol L-I) x volume (mL)

ofKHCsH404 1000
0.100mo1L -1 x 25.0mL
1000
= 2.5 X 10-3 mol
4. Calculate the number of moles of NaOH used to reach equivalence.

As indicated in point 2 above, I mole of NaOH is equivalent to
mole of KHCgH404. Therefore:
No. . of molesd = no. 0 f mo 1es 0

f' IKH C sH 04 m
. t hee trtran
ti d
4
o f NaOH use
= 2.5 X 10-3 mol
5. Determine the concentration of the NaOH solution:

Concentration of NaOH 1000 x amount of NaOH (mol)
solution (mol L -I) volume of NaOH solution used (mL)
1000 x 2.5 X 10-3 (mol)
23.50 (mL)
= 0.106molL-1

------~0 Complexometric titrations
Complexometric titrations are mainly used to determine the concentration of

cations in solution. The method is based on the competition between a metal
ion (for example) and two Iigands, one of which acts as an indicator and the
other is a component of a standard solution.
Some knowledge of the principles of metal-ligand binding is required in
order to understand this method.
Types of ligand
Ligands are chemical species that co-ordinate with metal ions to form a
complex. They are classified on the basis of the number of points of
attachment to the central ion.
• Monodentate ligand - here the ligand is bound to the central ion at only
one point, e.g. H20, NH3.
• Bidentate ligand - this has two points of attachment to the central ion,
e.g. ethylenediamine (en) (Fig. 23.1).
Fig. 23.1 Structure of [Co(enbl3+. It is a six-
co-ordinate octahedral complex of • Multidentate ligand - these have several points of attachment, e.g.
ethylenediamine (en) with cobalt (Ill).The ethylenediaminetetra-acetic acid (EDTA), which is a hexadentate ligand
complex has three five-membered rings. (six points of attachment) (Fig. 23.2).
HOOC- CHz, "CHz- COOH The basis of a complexometric titration involving EDTA
"N-CHz-CHz-N,
HOOC- CHz CHz - COOH The metal ion under investigation is bound to an indicator in solution (under
strict pH control). This solution is then titrated against a standard solution of
(a)
EDTA. This can be expressed in the form of an equation:
Metal-indicator + EDTA --+ metal-EDTA + indicator [23.1]
For example, if the indicator being used was solochrome black, the metal-
indicator solution would be red while the colour of the free indicator would
be blue (in the pH range 7-11). The reaction takes place if the EDTA
displaces the indicator from the metal-indicator complex. Therefore the
metal-EDT A complex must be more stable thermodynamically than the
metal-indicator complex.
Stability of complexes
The thermodynamic stability of a species is an indication of the extent to
(hI
which that species will be formed (under certain conditions and provided that
Fig.23.2 Structure of EDTA.(a) EDTAcontains it is allowed to reach equilibrium).
two donor N atoms and four donor 0 atoms. It As an example consider the general case of a metal, M, in solution
can therefore form a hexadentate complex together with a monodentate ligand, L. It is possible to describe this system
(b) with a metal ion, e.g. Pb2+.
in terms of step-wise equilibria:
M+L=ML K1 = [ML]/[M][L] [23.2]
ML+ L= ML2 K2 = [ML2]/[ML][L] [23.3]
Or, in general terms:
ML(I7_1)+ L = ML17 [23.4]
where Kt, K2,··· K17 are step-wise stability constants.

Complexometric titrations
An alternative approach for expressing the equilibria might be as follows:
M+L=ML Ih = [ML]/[M][L] [23.5]
M + L2 = ML2 f32 = [ML2]/[M][Lf [23.6]

Or, in general terms:
M+Ln=MLn f3n = [MLn]/[M][L]n [23.7]

where f31, f32, ... , f3n are the overall stability constants and are related to the
step-wise stability constants as follows:
[23.8]
Factors influencing the stability of complexes

The stability of a complex is related to the ability of the metal ion to complex
with a given ligand, and to the characteristics of the ligand.
End-points can be determined more easily when a single complex is formed
rather than when the complex is formed in a step-wise fashion. This can be
achieved by using the aminopolycarboxylic acid, EDTA (Fig. 23.2).
In equations, EDTA can be expressed as H4Y. The disodium salt Na2H2Y is
frequently used as a source of the complex-forming ion, H2v>. Thus the
typical reaction of EDT A with a metal ion can be written in the following form:
M2+ + H2y2- -> My2- + 2H+ [23.9]
Table 23.1 Stability constants of selected The reaction of a metal ion with EDT A is always in the ratio I: I. The
metal-EDTAcomplexes (expressed as log 1<)* stability constants of selected metal-EDTA complexes are given in Table
23.1.
Ion logk Ion logk
Mg2+ 8.7 Ni2+ 18.6 The detection of the end-point in titrations involving EDTA is most
Ca2+ 10.7 Cu2+ 18.8 commonly achieved using a metal-ion indicator, i.e. a compound that
Fe2+ 14.3 Hg2+ 21.9
C02+ Sc3+
changes its colour when it complexes with a particular metal ion. The
16.3 23.1
A13+ 16.3 Cr3+ 24.0 structures of selected metal-ion indicators are shown in Fig. 23.3 and the
Cd2+ 16.6 In3+ 24.9 properties of a variety of metal-ion indicators are given in Table 23.2.
*Ionicstrength of solution was 0.1 at 20°C.

Adapted from Voge/'s Textbook of Quantitative Types of complexometric titration
Inorganic Analysis, 4th Edn, J. Bassett, R.C.
Denney, G.H.Jeffery and J. Mendham, Longman Direct titration
Scientific and Technical, Harlow, (1978)p. 264. In this case, the metal ion is titrated with a standard solution of EDTA. The
solution containing the metal ion is buffered to an appropriate pH at which
the stability constant of the metal-EDTA complex is large. The free indicator
OH OH
has a different colour from that of the metal-indicator complex.
+Na-03s-t-N~N-OO
N02 Back titration
In certain circumstances a particular metal ion cannot be titrated directly.
Solochrome black
Ieriochromeblack T) This includes situations where:
• The metal ion precipitates in the absence of EDTA.

• The metal ion reacts too slowly with EDT A.
• The metal ion forms an inert complex.
• No suitable indicator is available.
Calmagite
In these cases a back titration is required. This involves addition of a known

Fig.23.3 Examples of metal-ion indicators:
solochrome black and calmagite. excess of EDTA to the metal ion (buffered to an appropriate pH). Then, the
excess EDT A is titrated with a standard solution of a different metal ion. The
choice of a second metal ion is important as it must not displace the analyte
metal ion from its EDT A complex.

Table 23.2 Properties of selected indicators
Murexide
< pH 9 (H4In-) Red-violet orange (Cu2+), yellow (Ni2+ and C02+)
pH9-11 (H3In2-) Violet and red (Ca2+)
> pH 11 (H2In3-) Blue
Solochrome black
< pH 5 (H2In-) Red In pH range 7-11 colour change is
pH7-11 (Hln2-) Blue blue-red (Mg, Mn, Zn, Cd, Hg, Pb, Cu,
> pH 11.5 (In3-) Orange AI, Fe, Ti, Co, Ni and Pt metals)
Calmagite
<pH 5 (H2In-) Red Same colour change as solochrome
pH 7-9 (Hln2-) Blue black but clearer and sharper
>pH 11.4 (In3-) Red-orange
Pyrocatechol violet
< pH 1.5 (H4In) Red In pH range 2-6, yellow to blue
pH 2-6 (H3In-) Yellow (Bi and Th); pH 7 violet to blue
pH 7 (H2In2-) Violet (Cu2+, Zn2+, Cd2+, Ni2+ and C02+)
>pH 10 (In4-) Blue
Practical considerations
• pH adjustment is critical in EDTA titrations. The pH is monitored with a
pH meter or pH test paper.
• The metal ion under investigation should ideally be approximately
0.25 mM in a volume of 50--150mL of solution. Dilution of the metal ion
may be necessary to avoid end-point detection problems.
• Do not add excess indicator, as too intense a colour can lead to
problems, e.g. masking of the colour change.
• It is sometimes difficult to detect the end-point because the colour change
can be slow to develop. Stirring is recommended to assist colour
transformation.
• The use of metal-ion indicators to indicate the end-point of complexometric
titrations is based on a specific colour change. Some individuals may find it
difficult to detect a particular colour change (e.g. those with colour
blindness). Alternative approaches for end-point detection are available
based on a colorimeter/spectrophotometer (devices for measuring colour,
see Chapter 26) or electrochemical detection (see Chapter 34).
Example calculation
A solution of Ni2+ (25.00mL) was titrated with 0.1036mol L -I EDTA at
pH 5 and required 20.25 mL for the metal-indicator to change colour. What
is the concentration (g L -1) of Ni2 +? The atomic weight of nickel is
58.71 g mol ":
the test substance
[23.10]
2. Determine the equivalences of the reacting species:
1 mole of EDTA is equivalent to 1 mole of Ni2+

3. Calculate the number of moles of standard substance (EDTA) used to

reach the end-point of the reaction:
moles of EDTA = volume (L) x molarity (mol L -I)
= 20.25 X 10-3 Lx 0.1036mol L-1
= 2.098 x 10-3 mol

4. Calculate the corresponding number of moles of Ni2+ present III the
25 mL of nickel solution.
Since 1 mole of EDTA is equivalent to 1 mole of Ni2+:
moles of Ni2+ = 2.098 X 10-3 mol
5. Calculate the concentration of the Ni2+ solution:

concentration of Ni2+ 1000 x 2.098 X 10-3 (mol)
solution (mol L -1) 25 (mL)
= 83.92 x 10-3 mol L-1
In this instance, the Ni2 + concentration is required in g L -I.
The molecular weight of nickel is 58.71 g mol:":
concentration of molecular weight (g mol ")
Ni2+ solution (g L -1) = x molarity (mol L -I)
= 58.71 x 83.92 x 10-3

= 4.927 gL-1

------~G
Redox titrations
All reduction-oxidation reactions involve a transfer of electrons. The

oxidizing agent accepts electrons, and the reducing agent donates electrons.
To establish the reaction for a redox titration it is necessary to determine the
'half-equation' for both the oxidizing agent and the reducing agent. By
adding the two 'half-equations' it is possible to determine the overall
equation for the titration. The basic theory of electrochemistry is described in
Chapter 34.
One of the most common oxidants is potassium permanganate which in
acidic solution can undergo the following reaction:
[24.1]
Unfortunately, potassium permanganate is not obtainable in high enough

purity and can undergo decomposition by exposure to sunlight. Therefore
it cannot be used as a primary standard (p. 143). However, it can be used
in redox titrations provided it is standardized with sodium oxalate (which
is available in high purity). The redox reaction involving oxalate is as
follows:
[24.2]
The overall reaction between permanganate and oxalate can be obtained by

balancing the electrons on each side of the equation. This can be achieved by
multiplying eqn [24.1] by 2 and eqn [24.2] by 5, and then combining them as
follows:
Another common method for the standardization of potassium perman-

ganate is to use iron (Il):
[24.4]
The combined equation is obtained by multiplying eqn [24.4] by 5 and adding

to eqn [24.1]:
[24.5]
Potassium permanganate has a major advantage when used for titrations in

that it can act as its own indicator.
A list of other common oxidizing agents and reducing agents is given in
Table 24.1.
Table 24.1 Common oxidizing and reducing agents used in redox titrations
Ce4+ Ceric AsO§- Arsenite

Cr20~- Dichromate Fe2+ Ferrous
H202 Hydrogen peroxide NH20H Hydroxylamine
1°3 Iodate Sn2+ Stannous
MnOi Permanganate S20~- Thiosulphate

Redox titrations
Example calculation
Standardization of potassium permanganate with a primary
standard, sodium oxalate
An accurately weighed portion of sodium oxalate (0.1550 g) was dissolved in
dilute sulphuric acid (250mL). Whilst maintaining the temperature of the
solution above 70 "C, it was titrated to equivalence with potassium perman-
ganate solution (18.5 mL). What is the molarity of potassium permanganate?
the test substance (using the two half-equations [24.1] and [24.2]):
2MnO; + 16H+ + 5CzO~- = 2Mnz+ + lOCOz + 8HzO [24.3]

2. Determine the equivalences of the reacting species:
2 moles of MnO; are equivalent to 5 moles of C20~- .
3. Calculate the number of moles of standard substance (sodium oxalate)

used to reach the end-point of the reaction.
The molecular weight of sodium oxalate is 134 g mol ": Therefore
0.1550 g of sodium oxalate is equivalent to:
0.1550(g) _ 1.157 x 10-3 mol

134 (g mol ")
4. Calculate the corresponding number of moles of potassium perman-
ganate present in the volume of titrant added.
From the equation:
5 moles of Na2CZ04 = 2 moles of KMn04

Therefore,
moles of KMn04 used in the titration = ~x 1.157 X 10-3 mol
= 4.628 x 10-4 mol
5. Calculate the concentration of the KMn04 solution:
Concentration of KMn04 1000 x amount of KMn04 (mol)
solution (molL-I) Volume of KMn04 solution used (mL)
1000 x 4.628 X 10-4 (mol)
18.5 (mL)
= 0.025 molL -I

------~0 Precipitation titrations
Precipitation is the term used to describe the process whereby a substance

leaves solution rapidly, forming either a crystalline solid or amorphous solid
(the precipitate). In the case of a precipitation titration, this process occurs
when the analyte forms a precipitate with the titrant. The most common
types of precipitation titrations use silver nitrate as the titrant. They are often
referred to as argentimetric titrations.
Titration curves used in precipitation reactions usually use a concentration-
dependent variable called the 'p function' rather than the concentration itself.
The p function for a species X is defined as follows:
pX = -loglO [X] [25.1]
For example, in the titration of 100mL of 0.1 mol L -I NaC1 with 0.1 mol L -I
AgN03 the initial concentration of [Cl"] is 0.1 mol L -I, so by using eqn [25.1]
the p function is 1 or pCl- = 1.
When 25mL of O.lmoIL-1 AgN03 has been added, 75mL of NaCl
remains in a total volume of 125mL. Therefore, the concentration of the
chloride ion is given by
[Cl"] = 75mL x 0.1 M _ 6 x 10-2 molL-1 [25.2]

125mL
and pCl- = 1.22. (Note that the solubility product, Kg, of AgCl is
Table 25. 7 Titration of 100 mL of 0.1 M NaCI 1.1 x 10-10, see p. 50.)
with 0.1 M AgN03. (Note that Ks. for
Therefore:
AgCI = 1.1 x 10-1°)
[25.3]
or
o 0.1 0.0
pAg+ + pCl- = 9.96 = pAgCl [25.4]
25 1.2 8.7
50 1.5 8.5
90 2.3 7.7 It was found above that pCl- = 1.22, hence pAg+ = 9.96 - 1.22 = 8.74. In a
95 2.6 7.4
similar manner, the pAg+ values can be calculated.
98 3.0 7.0
99 3.3 6.7 At the equivalence point [Ag"] = [Cl"], Therefore:
99.5 3.6 6.4
99.8
99.9
4.0
4.3
6.0
5.7
[Ag+] = [Cn = ..[J{; = vU X 10-10 = 1.05 X 10-5 [25.5]
100 5.0 5.0
100.1 5.7 4.3 pAg+ = -10g(1.05 x 10-5) = 4.98 [25.6]
100.2 6.0 4.0
100.5 6.4 3.6
101 6.7 3.3 Beyond the equivalence point the situation changes. For 100.1mL AgN03
102 7.0 3.0 solution:
105 7.4 2.6
110 7.7 2.3
120 8.0 2.0 [A+]=0.lmLxO.1M=5x10-5 lL-1 [25.7]
g 200.1 mL mo
130 8.1 1.9
140 8.2 1.8
150 8.3 1.7 or pAg+ = 4.3. Therefore,
Adapted from Vogef's Textbook of

pCl- = pAgCl - pAg+ = 9.96 - 4.3 = 5.66
Quantitative Inorganic Analysis, 4th Edn, J.
Bassett, R.C. Denney, G.H. Jeffery and
J. Mendham, Longman Scientific and Technical, Values calculated in this way up to the addition of 150mL of 0.1 M AgN03
Harlow (1978), p. 280. are given in Table 25.1 and the titration curve in Fig. 25.1.

Precipitation titrations
equivalence point, pAg+ = 5 End-point determination

10.0
Three techniques are commonly used to determine the end-point III
8.0
precipitation titrations. They are:
+ 6.0
:te, 1. potentiometric methods;
4.0 2. chemical indicator methods;
2.0 3. light-scatterring methods, exemplified by turbidimetry or nephelometry.
0.0 Only indicator methods will be discussed further. Three types of indicator
0 50 100 150 methods can be applied to determine the end-point of an Ag" and a Cl-
0.1 M AgN03 (mL)
titration. These are:
Fig. 25.1 Precipitation titration curve. Initial 1. Mohr titration, which involves the formation of a coloured
and final slopes are drawn (see Fig. 22.3) and a
precipitate by reaction with the indicator. For example, in the
parallel line is drawn such that the mid-point is
on the curve. This is the equivalence point. determination of chloride concentration with silver nitrate a small
amount of potassium chromate solution is added as an indicator.
This results in the formation of a red silver chromate (Ag2Cr04)
precipitate at the end-point:
2Ag+ + CrO~- --'> Ag2Cr04(s) [25.8]

(red)
In this case, the precipitate may form slightly after the end-point, but
this error can usually be neglected. Also, the titration should be done in
neutral or slighly alkaline solution (pH 6.5-9) otherwise silver chromate
might not be formed.
2. Volhard titration, which involves the formation of a soluble coloured
compound. This approach is exemplified by the quantitative analysis of
chlorides, bromides and iodides by back titration. In this case, the halide
is titrated with silver:
[25.9]
Excess silver ions are then titrated with standard potassium thiocyanate
solution in the presence of an iron (Ill) salt:
[25.10]
When all the Ag" has been reacted, the SCN- reacts with Fe3+ to form
a red complex, indicating the end-point:
Fe3+ + SCN- --'> FeSCN2+ [25.11]
(red)
A problem with the determination of chloride by this approach is that the

end-point coloration slowly fades, as AgCl is more soluble than AgSCN.
As a consequence the AgCl slowly dissolves to be replaced by the
FeSCN2+. Two approaches are possible to prevent this secondary reaction
from taking place. The most common method is to filter off the AgCl and
titrate only the Ag" left in solution. Alternatively, add a few millilitres of
an immiscible liquid (e.g. nitrobenzene) to the titrand prior to the back
titration. The nitrobenzene acts to 'coat' the AgCI precipitate, thereby
isolating it from the SCN-.
3. Fajans titration, which involves the adsorption of a coloured indicator
onto the precipitate at the end-point, resulting in a colour change.
During this adsorption process a change occurs in the indicator resulting
Fig. 25.2 Structure of dichlorofluorescein. in a change of colour. The indicators used for this are often anionic dyes,

Precipitation titrations
Table 25.2 Selected applications of e.g. fluorescein or eosin. The most common indicator for AgCl is
precipitation titrations
dichlorofluorescein (Fig. 25.2) (this is greenish yellow III solution but
changes colour to pink when it is ads or bed on AgCl).
Mohr method: A92Cr04 used as Selected examples of precipitation titrations are shown in Table 25.2.
end-point
Volhard method: precipitate
removal is unnecessary
CI-, CW, Volhard method: precipitate
CO~- removal is required
CI-, Br-, 1-, Fajans method: titration with
SCN- Aq '. Detection with fluorescein,
dichlorofluorescein and eosin
Titration with Th(N03)4 to
produce ThF4. End-point
detection with alizarin red S
Adapted from: Quantitative chemical analysis,

4th Edn, D.C. Harris, W.H. Freeman, New York
(1995), p. 176.

Resources
Resources for classical techniques
General books
Main supplementary text:
Mendham, J., Denney, R.e., Barnes, J.D. and Thomas, MJ.K. (2000)
Harlow, Essex.
Other useful sources (chronological order):

Bennett, S.W. and O'Neale, K. (1999) Progressive development of practical
skills in chemistry. A guide to early undergraduate experimental work, Royal
Crawford, K. and Heaton, A. (1999) Problem Solving in Analytical
Chemistry, Royal Society of Chemistry, Cambridge.
Day R.A. and Underwood, A.L. (1991) Quantitative Analysis, 6th Edn,
Prentice Hall, Harlow, Essex.
Rubinson, J.F. and Rubinson, KA. (1998) Contemporary Chemical Analysis,
Prentice Hall, Harlow, Essex.
Skoog, DA., West, D.M. and Holler, F.J. (1996) Fundamentals of Analytical
Chemistry, 7th Edn, Saunders College Publishing, Orlando, Florida.
Specific books on qualitative analysis

Main supplementary text:
Svehla, G. (1989) Vogel's Qualitative Inorganic Analysis, Longman, Harlow,
Essex.
Other useful sources (chronological order):
Whitten, K.W., Davis, R.E. and Peck, M.L. (2000) General Chemistry with
Qualitative Analysis, 6th Edn, Saunders College Publishing, Orlando, Florida.
Hardcastle, W.A. Qualitative Analysis. A guide to best practice, Royal Society
of Chemistry, Cambridge.
Videos
Basic Laboratory Skills, LGC, Royal Society of Chemistry, Cambridge (1998).
Further Laboratory Skills LGC, Royal Society of Chemistry, Cambridge
(1998).
CD-ROMs
Chemistry Video Consortium, Practical Laboratory Chemistry, Educational
Media Film and Video Ltd, Harrow, Essex, UK - Inorganic analysis
(gravimetric analysis).
Media Film and Video Ltd, Harrow, Essex, UK ~ Volumetric techniques
(using a balance, using a pipette, using a burette and making-up solutions).
Media Film and Video Ltd, Harrow, Essex, UK - Volumetric analyses
methods (doing a titration, some common end-points and potentiometric
titrations).

-0 Basic spectroscopy
Light is most strictly defined as that part of the spectrum of electromagnetic

radiation detected by the human eye. However, the term is also applied to
radiation just outside that visible range, e.g. ultraviolet (UV) and infrared
(IR) 'light'. Electromagnetic radiation is emitted by the sun and by other
sources (e.g. an incandescent lamp) and the electromagnetic spectrum is a
broad band of radiation, ranging from cosmic rays to radio waves (Fig. 26.1).
Most chemical experiments involve measurements within the UV, visible and
IR regions (generally, within the wavelength range 200-1000 nm).
Radiation has the characteristics of a particle and of a vibrating wave,
travelling in discrete particulate units, or 'packets', termed photons. A
quantum is the amount of energy contained within a single photon (it is
important not to confuse these two terms, although they are sometimes used
interchangeably in the literature). In some circumstances, it is appropriate to
measure light in terms of the number of photons, usually expressed directly in
moles (6.02 x 1023 photons = 1mol). Alternatively, the energy content
(power) may be measured (e.g. in Wm-2). Radiation also behaves as a
1 04 vibrating electrical and magnetic field moving in a particular direction, with
radio the magnetic and electrical components vibrating perpendicular to one
waves
another and perpendicular to the direction of travel. The wave nature of
1 01 radiation gives rise to the concepts of wavelength (A, usually measured in
nm), frequency (v, measured in S-I, but often recorded in hertz, Hz), speed
(c, the speed of electromagnetic radiation, which is 3 x 108 m S-l in a
1 00
vacuum), and direction. In other words, radiation is a vector quantity, where:
infrared microwaves
[26.1]
800 770 nm 1 0-1
Sometimes, it is necessary to rearrange the equation such that:
l
red E
-S
infrared c
700 1 0-4 -'=
0>
c; V=- [26.2]
ID
Q;
A
>
orange ~
yellow visible
1 0-6 Introduction to spectroscopy
7
green ultraviolet The absorption and emission of electromagnetic radiation of specific energy
1 0-8 (wavelength) is a characteristic feature of many molecules, involving the
blue
movement of electrons between different energy states, in accordance with the
violet x-rays laws of quantum mechanics. Electrons in atoms or molecules are distributed at
400 1 0-10 various energy levels, but are mainly at the lowest energy level, usually termed
ultraviolet 390 nm the ground state. When exposed to energy (e.g. from electromagnetic
gamma rays radiation), electrons may be excited to higher energy levels (excited states),
300 1 0-12
with the associated absorption of energy at specific wavelengths giving rise to
an absorption spectrum. One quantum of energy is absorbed for a single
cosmic rays
electron transition from the ground state to an excited state. On the other
hand, when an electron returns to its ground state, one quantum of energy is
Fig. 26.1 The electromagnetic spectrum.
released; this may be dissipated to the surrounding molecules (as heat) or may
give rise to an emission spectrum. The energy change (D.E) for an electron
moving between two energy states, El and E2, is given by the equation:
[26.3]
where h is the Planck constant (p. 72) and v is the frequency of the
electromagnetic radiation (expressed in Hz or s"). By substituting for
Instrumental techniques 163

Basic spectroscopy
frequency in [26.3J it IS possible to rearrange this equation to give the

expression:
!:::.E= he [26.4J
A
UV Ivisible spectrophotometry
This is a widely used technique for measuring the absorption of radiation in
the visible and DV regions of the spectrum. A spectrophotometer is an
instrument designed to allow precise measurement at a particular wavelength,
while a colorimeter is a simpler instrument, using filters to measure broader
wavebands (e.g. light in the green, red or blue regions of the visible
spectrum).
Principles of light absorption

Two fundamental principles govern the absorption of light passing through a
solution:
1. The absorption of light is exponentially related to the number of
molecules of the absorbing solute that are encountered, i.e. the solute
concentration [C].
2. The absorption of light is exponentially related to the length of the light
path through the absorbing solution, l.
These two principles are combined in the Beer-Lambert relationship, which is

usually expressed in terms of the intensity of the incident light (lo) and the
emergent light (l):
10glO(9) = 8[C]1 [26.5J
where 8 is a constant for the absorbing substance at the wavelength the

measurement is made and is termed the absorption coefficient or absorptivity,
[C] is expressed as either molL -1 or g L -I (see p. 45) and I is given in cm. This
relationship is extremely useful, since most spectrophotometers are constructed
to give a direct measurement of 10glO(IO/I), termed the absorbance (A), or
extinction (H), of a solution (older texts may use the outdated term optical
density). Note that for substances obeying the Beer-Lambert relationship, A is
linearly related to [C]. Absorbance at a particular wavelength is often shown as
a subscript, e.g. A550 represents the absorbance at 550 nm. The proportion of
light passing through the solution is known as the transmittance (T), and is
calculated as the ratio of the emergent and incident light intensities.
Some instruments have two scales:
1. An exponential scale from zero to infinity, measuring absorbance.
2. A linear scale from 0 to 100, measuring (per cent) transmittance.
sample
cell For most practical purposes, the Beer-Lambert relationship will apply and
crr-=[j'~U~[)4Io]~~I
light I detector amplifier/
you should use the absorbance scale.
u V/visible spectrophotometer
source exit test readout The principal components of a U'V/visible spectrophotometer are shown in
slit solution
Fig. 26.2. High-intensity tungsten bulbs are used as the light source in basic
Fig.26.2 Components of a UV/visible instruments, capable of operating in the visible region (i.e. 40Q-700nm).
spectrophotometer. Deuterium lamps are used for DV spectrophotometry (200-400 nm); these
164 Instrumental techniques

Basic spectroscopy
lamps are fitted with quartz envelopes, since glass does not transmit UV
radiation.
The spectrophotometer is a major improvement over the simple colorimeter
since it uses a diffraction grating to produce a parallel beam of monochromatic
light from the (polychromatic) light source. In practice the light emerging from
such a monochromator is not of a single wavelength, but is a narrow band of
wavelengths. This bandwidth is an important characteristic, since it determines
the wavelengths used in absorption measurements - the bandwidth of basic
spectrophotometers is around 5-10 nm while research instruments have
bandwidths of less than 1nm.
Bandwidth is affected by the width of the exit slit (the slit width), since the
bandwidth will be reduced by decreasing the slit width. To obtain accurate
data at a particular wavelength setting, the narrowest possible slit width
should be used. However, decreasing the slit width also reduces the amount
of light reaching the detector, decreasing the signal-to-noise ratio. The extent
to which the slit width can be reduced depends upon the sensitivity and
stability of the detection/amplification system and the presence of stray light.
Most Uv/visible spectrophotometers are designed to take cells (cuvettes)
with an optical path length of 10mm. Disposable plastic cells are suitable for
routine work in the visible range using aqueous and alcohol-based solvents,
while glass cells must be used for most other organic solvents. Glass cells are
manufactured to more exacting standards, so you should use optically
matched glass cells for accurate work, especially at low absorbances « 0.1),
where any differences in the optical properties of cells for reference and test
samples will be pronounced. Glass and plastic absorb UV light, so quartz
cells must be used at wavelengths below 300nm.
Before taking a measurement, make sure that cells are clean, unscratched,
dry on the outside, filled to the correct level and in the correct position in
their sample holders. Unwanted material can accumulate on the inside faces
of glass/quartz cells, so remove any deposits using acetone on a cotton bud,
or soak overnight in I mol L -1 nitric acid. Corrosive and hazardous solutions
must be used in cells with tightly fitting lids or Teflon@ stoppers, to prevent
damage to the instrument and to reduce the risk of accidental spillage.
Basic instruments use photocells similar to those used in simple
colorimeters or photodiode detectors. In many cases, a different photocell
must be used at wavelengths above and below 550-600nm, owing to
differences in the sensitivity of such detectors over the visible waveband. The
detectors used in more sophisticated instruments, give increased sensitivity
and stability when compared with photocells.
Digital displays are increasingly used in preference to needle-type meters,
as they are not prone to parallax errors and misreading of the absorbance
scale. Some digital instruments can be calibrated to give a direct readout of
the concentration of the test substance.
Types of U'Vlvisible spectrophotometer

Basic instruments are single-beam spectrophotometers in which there is only
one light path. The instrument is set to zero absorbance using a blank
solution, which is then replaced by the test solution, to obtain an absorbance
reading. An alternative approach is used in double-beam spectrophotometers,
where the light beam from the monochromator is split into two separate
beams, one beam passing through the test solution and the other through a
reference blank. Absorbance is then measured by an electronic circuit which
compares the outputs from the reference (blank) and sample cells. Double-

Basic spectroscopy
beam spectrophotometry reduces measurement errors caused by fluctuations

in output from the light source or changes in the sensitivity of the detection
system, since reference and test solutions are measured at the same time
(Box 26.1). Recording spectrophotometers are double-beam instruments,
designed for use with a chart recorder or computer, either by recording the
difference in absorbance between reference and test solutions across a
predetermined waveband to give an absorption spectrum, or by recording the
change in absorbance at a particular wavelength as a function of time (e.g. in
a kinetic determination.
Quantitative spectrophotometric analysis

A single (purified) substance in solution can be quantified using the Beer-
Lambert relationship (eqn [26.5]), provided its absorptivity is known at a
particular wavelength (usually at the absorption maximum for the substance,
since this will give the greatest sensitivity). The molar absorptivity is the
absorbance given by a solution with a concentration of 1mol L -1

Basic spectroscopy
(= 1 kmol m -3) of the compound in a light path of 1cm. The appropriate

value may be available from tabulated spectral data (e.g. Anon., 1963), or it
can be determined experimentally by measuring the absorbance of known
concentrations of the substance (Box 26.1) and plotting a standard curve.
This should confirm that the relationship is linear over the desired
concentration range and the slope of the line will give the molar absorptivity.
Fluorescence
With most molecules, after electrons are raised to a higher energy level by
second absorption of electromagnetic radiation, they soon fall back to the ground
, exited state
.~~ : state by radiation1ess transfer of energy (heat) to the solvent. However, with
~~:
§ ~ r
some molecules, the events shown in Fig. 26.3 may occur, i.e. electrons may
c-, lose only part of their energy by non-radiant routes and the rest may be
first
exited state emitted as electromagnetic radiation, a phenomenon known as fluorescence.
Since not all of the energy that was absorbed is emitted (due to non-radiant
loss), the wavelength of the fluorescent light is longer than the absorbed light
(longer wavelength = lower energy). Thus, a fluorescent molecule has both
an absorption spectrum and an emission spectrum.
ground state Fluorescence spectrophotometry
Fig. 26.3 Energy levels and energy transitions The principal components of a fluorescence spectrophotometer (fluorimeter)
in fluorescence. are shown in Fig. 26.4. The instrument contains two monochromators, one
to select the excitation wavelength and the other to monitor the light emitted,
usually at 90° to the incident beam (though light is actually emitted in all
directions). As an example, the wavelengths used to measure the highly
G light source
fluorescent compound naphthalene are 270 nm (excitation) and 340 nm
(emission). Some examples of molecules with intrinsic fluorescence are given
in Table 26.1.
§ \ excitation monochromator
Table 26.1 Examples of compounds with intrinsic fluorescence
or filter
Drugs Aspirin, morphine, barbiturates, propanalol, ampicillin, tetracyclines

sample cell Vitamins Riboflavin, vitamins A, 86 and E, nicotinamide
\) (test solution) Pollutants Naphthalene, anthracene, benzopyrene
s: or filter
•
Compared with Uv/visible spectrophotometry,
has certain advantages, including:
fluorescence spectroscopy
Enhanced sensitivity (up to IOOO-fold),since the emitted light is detected

against a background of zero, in contrast to spectrophotometry where small
detector
changes in signal are measured against a large 'background' (see eqn [26.5]).
• Increased specificity, because not one, but two, specific wavelengths are
required for a particular compound.
@Iill]
readout
However, there are also certain drawbacks:
Fig. 26.4 Components of a fluorimeter • Not all compounds show intrinsic fluorescence, limiting its application.
(fluorescence spectrophotometer). Note that However, some non-fluorescent compounds may be coupled to fluorescent
sample cells for fluorimetry must have clear dyes, or fluorophores (e.g. alcohol ethoxylates may be coupled to
sides all round.
naphthoyl chloride).
• The light emitted can be less than expected owing to quenching, i.e. when
substances in the sample (e.g. oxygen) either interfere with energy
transfer, or absorb the emitted light (in some instances, the sample
molecules may self-quench if they are present at high concentration).

Basic spectroscopy
The sensitivity of fluorescence has made it invaluable in techniques in which

specific chemicals, e.g. polycyclic aromatic hydrocarbons and alcohol ethoxy-
lates, are linked to a fluorescent dye for detection in high-performance liquid
chromatography (p. 218).
Phosphorescence and luminescence

A phenomenon related to fluorescence is phosphorescence, which is the
emission of light following intersystem crossing between electron orbitals (e.g.
between excited singlet and triplet states). Light emission in phosphorescence
usually continues after the exciting energy is no longer applied and, since
more energy is lost in intersystem crossing, the emission wavelengths are
generally longer than with fluorescence. Phosphorescence has limited appli-
cations in chemical sciences.
Luminescence (or chemiluminescence) is another phenomenon in which
light is emitted, but here the energy for the initial excitation of electrons is
provided by a chemical reaction rather than by electromagnetic radiation. An
example is the action of the enzyme luciferase, extracted from fireflies, which
catalyses the following reaction:
luciferin + ATP + O2 ---+ oxyluciferin + AMP + PPj + CO2 + light [26.6]

The light produced is either yellow-green (560nm) or red (620nm). This
system can be used in biomolecular analysis of ATP, e.g. to determine ATP
concentration in a biological sample. Measurement can be performed using
the photomultiplier tubes of a scintillation counter (p. 237) to detect the
emitted light, with calibration of the output using a series of standards of
known ATP content.
Atomic spectroscopy
nebulizer Atoms of certain metals will absorb and emit radiation of specific wavelengths
flame when heated in a flame, in direct proportion to the number of atoms present.
r n rL_~
compressed ~ detector
air
Atomic spectrophotometric techniques measure the absorption or emission of
~ ~U~~'-/~ particular wavelengths of DV and visible light, to identify and quantify such
filter or amplifier / metals.
rnonochrornator readout
test burner Flame atomic emission spectrophotometry (or flame photometry)
solution
The principal components of a flame photometer are shown in Fig. 26.5. A
Fig. 26.5 Components of a flame photometer. liquid sample is converted into an aerosol in a nebulizer (atomizer) before
being introduced into the flame, where a small proportion (typically less than
I in 10000) of the atoms will be raised to a higher energy level, releasing this
energy as light of a specific wavelength, which is passed through a filter to a
photocell detector. Flame photometry can be used to measure the alkali
metal ions K+, Na" and Ca2+ in, for example, biological fluids and water
samples (Box 26.2).
Atomic absorption spectroscopy

This technique is applicable to a broad range of metal ions, including
those of Pb, Cu, Zn, etc. It relies on the absorption of light of a specific
wavelength by atoms dispersed in a flame. The appropriate wavelength is
provided by a hollow cathode lamp, coated with the element to be
analysed, focused through the flame and onto the detector. When the
sample is introduced into the flame, it will decrease the light detected in

Basic spectroscopy
direct proportion to the amount of metal present. Practical advantages

over flame photometry include:
• improved sensitivity;
• increased precision;
• decreased interference.
The technique can be used with or without a flame. In the flameless
technique several variations are possible, including a graphite furnace or cold
vapour, all of which are more sensitive than flame photometry. Further
details on atomic absorption spectroscopy are given in Chapter 27.

------~0 Atomic speetroscopy
Atomic spectroscopy is a quantitative technique used for the determination of

metals in samples. Atomic spectroscopy is characterized by two main
techniques: atomic absorption spectroscopy and atomic emission
spectroscopy. Atomic absorption spectroscopy (AAS) is normally carried out
with a flame (FAAS), although other devices can be used. Atomic emission
spectroscopy (AES) is typified by the use of a flame photometer (p. 168) or
an inductively coupled plasma. The flame photometer is normally used for
elements in groups I and 11 of the Periodic Table only, i.e. alkali and alkali
earth metals.
In both AAS and AES the substance to be analysed must be in solution. In
order to do quantitative analysis, i.e. determine how much of the metal is
present, the preparation of analytical standard solutions is necessary. While
the concentration range over which the technique can be used may be
different, for various instruments, the principles associated with the
preparation of analytical standard solutions are the same (Boxes 27.1-27.5).
Atomic Absorption Spectroscopy

The components of an atomic absorption spectrometer are a radiation
source, an atomization cell, a sample introduction system, a method of
wavelength selection and a detector (Fig. 27.2).
Radiation source
The main radiation source for AAS is the hollow-cathode lamp (HCL). The
HCL (Fig. 27.3) emits radiation characteristic of a particular element. The
choice of HCL for AAS is simple. For example, if you are analysing for lead,
you will need a lead-coated HCL. It is normal to pre-warm the HCL for
about 10min prior to use. This can be done either by using a separate pre-

Atomic spectroscopy
heater unit, capable of warming up several HCLs simultaneously, or by

inserting the HCL in the AAS instrument and switching on the current. The
lamp is typically operated at an electric current between 2 and 30 mA.
Atomization cell
Several types of atomization cell are available: flame, graphite furnace,
hydride generation and cold vapour. Flame is the most common. In the pre-
mixed laminar flame, the fuel and oxidant gases are mixed before they enter
the burner (the ignition site) in an expansion chamber. The more commonly
used flame in FAAS is the air-acetylene flame (temperature, 2500 K), while
the nitrous oxide-acetylene flame (temperature, 3150 K) is used for refractory
elements, e.g. AI. Both are formed in a slot burner positioned in the light
path of the HCL (Fig. 27.4).
In the graphite furnace atomizer, a small volume of sample (5-100,uL) is
introduced onto the inner surface of a graphite tube (or onto a platform
placed within the tube) through a small opening (Fig. 27.5). The graphite
tube is arranged so that light from the HCL passes directly through the
centre. Passing an electric current through the tube allows the operator to
program a heating cycle, with several stages (Fig. 27.6) including the
elimination of water from the sample (drying), removal of the sample matrix
(ashing), atomization of the analyte (analysis), and removal of extraneous
material (cleaning). An internal gas flow of inert gas (N2 or Ar) during the
drying and ashing stages removes any extraneous material.
Hydride generation is a sample introduction technique exclusively for
elements that form volatile hydrides (e.g. As, Se, Sn). An acidified sample
solution is reacted with sodium borohydride solution, liberating the gaseous
hydride in a gas-liquid separator. The generated hydride is then transported to

Atomic spectroscopy
an atomization cell using a carrier gas. The atomization cell is normally an

electrically heated or flame-heated quartz tube. Using arsenic as an example it is
possible to write the following equation for the generation of arsine (AsH3):
[27.1]
Cold vapour generation is the term exclusively reserved for mercury.

Mercury in a sample is reduced to elemental mercury by tin (ll) chloride
(eqn 27.2):
[27.2]
and the mercury vapour produced is transported to an atomization cell by a

carrier gas. The atomization cell consists of a long-path glass absorption cell
located in the path of the HCL. Mercury is monitored at a wavelength of
253.7 nm.
Sample introduction into the flame

Samples are almost exclusively introduced into flames as liquids. Solid
samples need to be converted to aqueous solutions using methods such as
decomposition (see p. 177). Once in the aqueous form the sample is
introduced into the flame using a nebulizer/expansion chamber.
The pneumatic concentric nebulizer (see also p. 176) consists of a stainless
steel tube through which a Pt/Ir capillary tube is located. The aqueous

Atomic spectroscopy
sample is drawn up through the capillary tube by the action of the oxidant
gas (air) escaping through the exit orifice that exists between the outside of
the capillary tube and the inside of the stainless steel tube. The action of the
escaping air and aqueous sample is sufficient to form a coarse aerosol in a
process termed the Venturi effect. The typical uptake rate of the nebulizer is
between 3 and 6mLmin-1.

Atomic spectroscopy
The expansion chamber (Fig. 27.7) has two functions. The first IS
concerned with aerosol generation the objectives of which are:
• to convert the aqueous sample solution into a coarse aerosol using the
oxidant gas;
• to disperse the coarse aerosol further into a fine aerosol, by interaction
with baffles located within the chamber;
• to condense any residual aerosol particles, which then go to waste.
The second function involves the safe pre-mixing of the oxidant and fuel
gases before they are introduced into the laminar flow burner.
Wavelength selection and detection

Fig.27.2 Components of an atomic absorption As AAS is used to monitor one metal at a time, the spectrometer used is
spectrometer. termed a monochromator. Two optical arrangements are possible; single and
double beam. The latter is preferred as it corrects for fluctuations in the HCL
hollow cathode
caused by warm-up, drift and source noise, thus leading to improved
connecting pins glass envelope
precision in the absorbance measurement. A schematic diagram of the optical
silica window
arrangement is shown in Fig. 27.8. The attenuation of the HCL radiation by
~
the atomic vapour is detected by a photomultiplier tube (PMT), a device for
proportionally converting photons of light to electric current.
Background correction methods

anode Ne or Ar
One of the main practical problems with the use of AAS is the occurrence of
Fig. 27.3 Components of a hollow-cathode molecular species that coincide with the atomic signal. One approach to
lamp (HCl). remove this molecular absorbance is by the use of background correction
methods. Several approaches are possible, but the most common is based on
the use of a continuum source, D2. In the atomization cell (e.g. flame)
absorption is possible from both atomic species and from molecular species
(unwanted interference). By measuring the absorption that occurs from the
radiation source (HCL) and comparing it with the absorbance that occurs
from the continuum source (D2) a corrected absorption signal can be
obtained. This is because the atomic species of interest absorb the specific
radiation associated with the HCL source, whereas the absorption of
radiation by the continuum source for the same atomic species will be
negligible.
Interferences in the flame

Interferences in the flame can be classified into four categories: chemical,
ionization, physical and spectral.
Chemical interferences occur when the analyte forms a thermally stable
Fig.27.4 Components of a slot burner for compound with a molecular or ionic species present in the sample solution.
FAAP. Examples include the suppression of alkaline earth metals due to the presence
of phosphate, silicate or aluminate in the sample solution in the air-acetylene
I
sample injection port
graphite tube
flame. The most well-known example of this is the absorption signal
suppression that occurs for Ca at 422.7 nm owing to increasing amounts of
phosphate. This signal suppression is due to the formation of calcium
f~~:~~~-+-(_)~ O ~
pyrophosphate, a thermally stable compound in the flame.
Ionization interferences occur most commonly for alkali and alkaline earth
tor electrical heating

metals. The low ionization potential of these metals can lead to their
ionization in the relatively hot environment of the flame. If this occurs, no
absorption signal is detected, since FAAS is a technique for measuring atoms
Fig. 27.5 Schematic diagram of a graphite not ions. This process can be prevented by the addition of an ionization
furnace atomizer. suppressor or 'buffer', e.g. an alkali metal such as Cs. Addition of excess Cs

Atomic spectroscopy
leads to its ionization in the flame in preference to the metal of interest, e.g.
Na. This process is termed the 'mass action' effect.
Physical interferences are due to the effects of the sample solution on
aerosol formation within the spray chamber. The formation of an aerosol is
dependent upon the surface tension, density and viscosity of the sample
solution. This type of interference can be controlled by the matrix matching
of sample and standard solutions, i.e. add the same sample components to
the standard solution, but without the metal of interest. If this is not possible,
it is then necessary to use the method of standard additions (Box 27.3).
Fig. 27.6 Heating cycle for a graphite furnace Spectral interferences are uncommon in AAS owing to the selectivity of
atomizer. 1. drying; 2. ashing; 3. analysis; 4. the technique. However, some interferences may occur, e.g. the resonance
cleaning and 5. cooling line of Cu occurs at 324.754 nm and has a line coincidence from Eu at
324.753 nm. Unless the Eu is 1000 times in excess, however, it is unlikely
fine aerosol to cause any problems for Cu determination. In addition to atomic
to burner spectral overlap, molecular band absorption can cause problems, e.g.
calcium hydroxide has an absorption band on the Ba wavelength of
553.55 nm while Pb at 217.0 nm has molecular absorption from NaCl.
Molecular band absorption can be corrected for using background
expansion
~ or cloud
correction techniques (see p. 174). The operation of a flame atomic
sample chamber absorption spectrometer is described in Box 27.6.
Atomic Emission Spectroscopy

water trap The main components of an atomic emISSIOn spectrometer are an
Fig.27.7 Schematic diagram of a nebulizer-
atomization and ionization cell, a method of sample introduction, the
expansion chamber for FAAS. spectrometer and detector. In contrast to AAS, no radiation source is
required.
Flame photometry (see also p. 168) is almost exclusively used for the
determination of alkali metals because of their low excitation potential (e.g.
Czerny-Turner sodium 5.14eV and potassium 4.34eV). This simplifies the instrumentation
monochromator
required and allows a cooler flame (air-propane, air-butane or air-natural
gas) to be used in conjunction with a simpler spectrometer (interference
1----~-----~ filter). The use of an interference filter allows a large excess of light to be
photomultiplier
,l, .;. tube viewed by the detector. Thus, the expensive photomultiplier tube is not
HCL required and a cheaper detector can be used, e.g. a photodiode or
: atom cell '\ :
-, PC read out
chopper
halt-silvered
r mirror
photoemissive detector. The sample is introduced using a pneumatic nebulizer
as described for F AAS (p. 172). Flame photometry is therefore a simple,
robust and inexpensive technique for the determination of potassium
Fig. 27.8 Schematic diagram of the optical
(766.5 nm) or sodium (589.0 nm) in clinical or environmental samples. The
arrangement for AAS. technique suffers from the same type of interferences as in F AAS. The
operation of a flame photometer is described in Box 26.2.
Inductively coupled plasma

A radio frequency inductively coupled plasma (ICP) is formed within the
confines of three concentric glass tubes or plasma torch (Fig. 27.9). Each
concentric glass tube has a tangentially arranged entry point through which
argon gas enters the intermediate (plasma) and external (coolant) tubes. The
inner tube consists of a capillary tube through which the aerosol is
introduced from the sample introduction system. Located around the plasma
torch is a coil of water-cooled copper tubing. Power input to the ICl? is
achieved through this copper, load or induction coil, typically in the range
0.5-1.5 kW at a frequency of27 or 40MHz.

Atomic spectroscopy
Initiation of the plasma is achieved as follows. The carrier gas flow is first
switched off and a spark added momentarily from a Tesla coil (attached to
the outer edge of the plasma torch). The spark, a source of 'seed' electrons,
causes ionization of the argon gas. The co-existence of argon, argon ions and
electrons constitutes a plasma located within the confines of the plasma torch
but protruding from the top in the shape of a bright white luminous bullet.
In order to introduce the sample aerosol into the ICP (7000-10 000 K) the
carrier gas is switched on and punches a hole into the centre of the plasma
creating the characteristic doughnut or toroidal shape. The emitted radiation
is viewed laterally (side-on) through the luminous plasma.
Sample introduction
The most common method of liquid sample introduction in ICP-AES is the
o o load coil nebulizer. The nebulizer operates in the same manner as that used for F AAS
o o but there are differences in its construction material and manufactured
tolerance (the nebulizer for ICP-AES generates a finer aerosol, but is more
injector tube inefficient). The pneumatic nebulizer consists of a concentric glass tube
through which a capillary tube passes (Fig. 27.10). The sample is drawn up
through the capillary by the action of the argon carrier gas escaping through
the exit orifice that exists between the outside of the capillary tube and the
inside of the glass concentric tube. The typical uptake rate of the nebulizer is
between 0.5 and 4mLmin-l. In common with FAAS, a means to reduce the
coarse aerosol generated to a fine aerosol is required. In ICP-AES
intermediate gas flow
terminology this device is called a spray chamber (Fig. 27.11).
outer gas flow

Spectrometers
The nature of the ICP is such that all elemental information from the sample is
contained within it. The only limitation is whether it is possible to observe all
the elemental information at the same time or one element at once. This
sample aerosol
limitation is associated not with the ICP but with the type of spectrometer used
Fig. 27.9 Schematic diagram of an inductively to view the emitted radiation. A monochromator allows measurement of one
coupled plasma. wavelength, corresponding to one element at a time, while a polychromator
liquid
sample
r \
argon gas
exit
orifice
allows multiwavelength or multielement detection. The former can perform
sequential multielement analysis, while the latter carries out simultaneous
multielement analysis. The typical wavelength coverage required for a
spectrometer is between 167nm (AI) and 852 nm (Cs).
Detectors
The most common detector for AES is the photomultiplier tube (see p. 174).
Fig. 27.10 Schematic diagram of a pneumatic An alternative approach for the detection of multielement (multiwavelength)
nebulizer. information is the charged-coupled device (CCD). A CCD is essentially an
array of closely spaced metal-insulator-semiconductor diodes formed on a
aerosol
wafer of semiconductor material. Incident light striking the CCD is converted
nebulizer----.-J ~ into an electrical signal.
~~ Interferences in ICP-AES
waste
Interferences for AES can be classified into two main categories, spectral and
Fig. 27.11 Schematic diagram of a spray matrix interferences. Spectral interference can occur as a result of an
chamber. interfering emission line from either another element or the argon source gas,
impurities within or entrained into the source, e.g. molecular species such as
N2. Such interferences can be eliminated or reduced either by increasing the
resolution of the spectrometer or by selecting an alternative spectral emission
line.

Atomic spectroscopy
Matrix interferences are often associated with the sample introduction

process. For example, pneumatic nebulization can be affected by the
dissolved-solids content of the aqueous sample, which affects the uptake rate
of the nebulizer and hence the sensitivity of the assay. Matrix effects in the
plasma source typically involve the presence of easily ionizable elements
(EIEs), e.g. alkali metals, within the plasma source.
Decomposition techniques for solid inorganic samples

Conversion of a solid matrix into a liquid matrix involves the decomposition
of the sample. One of the major problems in preparing solid samples for
trace element analysis is the potential risk of contamination. Contamination
can arise from several sources: the grade of reagents used; the vessels used for
digestion and the subsequent dilution of the sample; and human involvement.
In order to minimize the risk of contamination you should take the
following measures:
• Use the highest purity of reagents and acids, including the water used for
sample dilution.
• Use sample blanks in the analytical procedure, to identify the base level
of impurity in the reagents.

Atomic spectroscopy
• Soak sample vessels in an acid leaching bath (e.g. 10% v]» nitric acid) for
at least 24 hours, followed by rinsing in copious amounts of ultrapure
water.
• Store cleaned volumetric flasks with their stoppers inserted; cover beakers
with Clingfilm'P or store upside down to protect from dust.
• In addition to the wearing of a laboratory coat and safety glasses, it may
be necessary to wear 'contaminant'-free gloves and a close-fitting hat.
Decomposition involves the liberation of the analyte (metal) of interest from

an interfering matrix using a reagent (mineral/oxidizing acids or fusion flux)
and/or heat. An important aspect in the decomposition of an unknown
sample is the sample size (Box 27.4). You need to consider two aspects.
Firstly, the dilution factor required to convert the solid sample to an aqueous
solution (Box 27.5), and, secondly, the sensitivity of the analytical instrument,
e.g. FAAS.
Acid digestion
This involves the use of mineral or oxidizing acids and an external heat source
to decompose the sample matrix. The choice of an individual acid or
combination of acids depends upon the nature of the matrix to be decomposed.
For example, the digestion of a matrix containing silica, Si02 (e.g. a geological
sample), requires the use of hydrofluoric acid (HF). A summary of the most
common acids used for digestion and their application is shown in Table 27.3.
Once you have chosen an appropriate acid, place your sample into an
appropriate vessel for the decomposition stage. Typical vessels include an open
glass beaker or boiling tube for conventional heating or for microwave heating,
a PTFE or Teflon® PFA (perfluoroalkoxyvinylether) vessel. A typical micro-
wave system operates at 2.45 GHz with up to 14 sample vessels arranged on a
rotating carousel; commercial systems have additional features such as: a
PTFE-lined cavity; a safety vent (if the pressure inside a vessel is excessive the
vent will open, allowing the contents to go to waste); and an ability to measure
both the temperature and pressure inside the digestion vessels. The procedure
for acid digestion of a sample is shown in Box 27.7.
Table 27. 3 Common acids* used for digestion
Hydrochloric acid (HCIl 110 Useful for salts of carbonates, some oxides and some sulphides. A weak reducing
agent; not generally used to dissolve organic matter
Hydrofluoric acid (HF) 112 For digestion of silica-based materials only. Cannot be used with glass containers
(use plasticware). In addition to laboratory coat and safety glasses, extra safety
precautions are needed, e.g. gloves. In case of spillages, calcium gluconate gel is
required for treatment of skin contact sites and should be available during use;
evacuate to hospital immediately if skin is exposed to liquid HF
Nitric acid (HN03) 122 Useful for the digestion of metals, alloys and biological samples. Oxidizing attack
on many samples not dissolved by HCI; liberates trace metals as the soluble
nitrate sa It
338 Useful for releasing a volatile product; good oxidizing properties for ores, metals,
alloys, oxides and hydroxides. Often used in combination with HN03. Note:
Sulphuric acid must never be used in PTFE vessels (melting point 327 DC)
Hydrochloric/nitric acids A 3: 1 v/v mixture of HCI and HN03 is called aqua regia. It forms a reactive
(HCI/HN03) intermediate, NOel. Useful for digesting metals, alloys, sulphides and other ores
*AII concentrated acids should be used only in a fume cupboard.

Atomic spectroscopy
Other methods of sample decomposition

The use of acid(s) and heat is probably the most common approach to the
decomposition of samples. However, several alternatives exist including dry
ashing and fusion.
Dry ashing involves heating the sample in air in a muffle furnace at 400-
800 QC to destroy the sample matrix, e.g. soil. After decomposition, the
sample residue is dissolved in acid and quantitatively transferred to a
volumetric flask prior to analysis. The method may lead to the loss of volatile
elements, e.g. Hg, As.
Some substances, such as silicates and oxides, are not always destroyed by
the direct action of acid and heat. In these situations an alternative approach
is required. Fusion involves the addition of a lO-fold excess of a suitable
reagent (e.g. lithium metaborate or tetraborate) to a finely ground sample.
The mixture is placed in a metal crucible, e.g. Pt, and then heated in a muffle
furnace at 90~ 1000 "C. After heating (from several minutes to several hours)
a clear 'melt' should result, indicating completeness of the decomposition.
After cooling, the melt is dissolved in HF (Table 27.3). This process can lead
to a higher risk of contamination.

-------0 Infrared spectroscopy
In addition to ultraviolet-visible (UV-vis) spectroscopy (p. 164), there are

three other essential techniques that you will encounter during your laboratory
course. They are:
1. Infrared (IR) spectroscopy: this is concerned with the energy changes
involved in the stretching and bending of covalent bonds in molecules.
2. Nuclear magnetic resonance (NM R) spectroscopy: this involves the absorp-
tion of energy by specific atomic nuclei in magnetic fields and is probably
the most powerful tool available for the structural determination of
molecules (Chapter 29).
3. Mass spectrometry (MS): this is based on the fragmentation of
compounds into smaller units. The resulting positive ions are then
separated according to their mass-ta-charge ratio (mjz) (Chapter 30).
As with Uv-vis spectroscopy, IR and NMR spectroscopy are based on the
interaction of electromagnetic radiation with molecules, whereas MS is
different in that it relies on high-energy particles (electrons or ions) to break
up the molecules. The relationship between the various types of spectroscopy
and the electromagnetic spectrum is shown in Table 28.1.
Infrared spectroscopy
A covalent bond between two atoms can be crudely modelled as a spring
connecting two masses and the frequency of vibration of the spring is defined
by Hooke's law (eqn [28.1]), which relates the frequency of the vibration (v)
to the strength of the spring, expressed as the force constant (k), and to the
masses (m] and m2) on the ends of the spring (defined as the reduced mass
i1 = (m) x m2) -;.-( m) + m2)).
[28.1]
V=2~~
In simple terms, this means that:

• the stretching vibration of a bond between two atoms will increase in
frequency (energy) if on changing from a single bond to a double bond
and then to a triple bond between the same two atoms (masses), i.e. the
spring gets stronger. For example,
Table 28.1 The electromagnetic spectrum and types of spectroscopy
y-rays Atomic nuclei < 0.1 nm y-ray spectroscopy

X-rays Inner shell electrons 0.01-2.0nm X-ray fluorescence (XRF)
Ultraviolet (UV) Ionization 2.0-200 nm Vacuum UV spectroscopy
UV/Visible Valency electrons 200-800nm UV/visible spectroscopy
Infrared Molecular vibrations 0.8-300llm IR and Raman spectroscopy
Microwaves Molecular rotations 1 mm to 30cm Microwave spectroscopy
Electron spin Electron spin resonance (ESR)
Radio waves Nuclear spin 0.6-10 m Nuclear magnetic resonance (NMR)

v for C=C > v for C=C > v for C-C

• as the masses of the atoms on a bond increases, the frequency of the
vibration decreases, i.e the effect of reducing the magnitude of fl; for
example,
v for C-H > v for C-C; v for C-H > v for C-D; v for O-H > v for S-H
Bonds can also bend, but this movement requires less energy than
stretching and thus the bending frequency of a bond is always lower than the
corresponding stretching frequency. When IR radiation of the same
frequency as the bond interacts with the bond it is absorbed and increases the
amplitude of vibration of the bond. This absorption is detected by the IR
spectrometer and results in a peak in the spectrum. For a vibration to be
detected in the IR region the bond must undergo a change in dipole moment
when the vibration occurs. Bonds with the greatest change in dipole moment
during vibration show the most intense absorption, e.g. C=O and c-o.
Since bonds between specific atoms have particular frequencies of
vibration, IR spectroscopy provides a means of identifying the type of bonds
in a molecule, e.g. all alcohols will have an O-H stretching frequency and all
compounds containing a carbonyl group will have a c=o stretching
frequency. This property, which does not rely on chemical tests, is extremely
useful in diagnosing the functional groups within a covalent molecule.
IR spectra
A typical IR spectrum IS shown III Fig. 28.1 and you should note the
following points:
'loT
100.00
0.00
4000 3500 3000 2500 2000 1500 1000 600 crn'
Fig. 28.1 IR spectrum of ethyl ethanoate CH3COOCH2CH3 as a liquid film.
• The x-axis, the wavelength of the radiation, is given in wavenumbers (v)

and expressed in reciprocal centimetres (cm"). You may still see some
spectra from old instruments using microns (/l, equivalent to the SI unit
'micrometres', pss», at 1 x 10-6 m) for wavelength; the conversion is given
by eqn [28.2]:
wavenumber (cm ") = ljwavelength (cm) = lOOOOjwavelength (/lm)

[28.2]

• The y-axis, expressing the amount of radiation absorbed by the molecule,

is usually shown as % transmittance (p. 164). When no radiation is
absorbed (all is transmitted through the sample) we have 100%
transmittance and 0% transmittance implies all radiation is absorbed at a
particular wavenumber. Since the y-axis scale goes from 0 to 100%
transmittance, the absorption peaks are displayed down from the 100%
line; this is opposite to most other common spectra.
• The cells holding the sample usually display imperfections and are not
completely transparent to IR radiation, even when empty. Therefore the
base line of the spectrum is rarely set on 100% transmittance and
quantitative applications of IR spectroscopy are more complex than for
IN-vis (p. 166).
motor 1l? spectror.neters

There are two general types:
1. Double-beam or dispersive instruments in which the IR radiation from a
single source is split into two identical beams. One beam passes through
the sample and the other is used as a reference and passes through air or
the pure solvent used to dissolve the sample. The difference in intensity
of the two beams is detected and recorded as a peak; the principal
Fig. 28.2 Schematic diagram of a double- components of this type of instrument are shown in Fig. 28.2. The
beam IR spectrometer. important controls on the spectrometer are:
(a) scan speed: this is the rate at which the chart moves - slower for
greater accuracy and sharp peaks;
(b) wavelength range: the full spectrum or a part of the IR range may
be selected;
(c) 100% control: this is used to set the pen at the 100% transmittance
line when no sample is present the base line. It is usual practice to
set the pen at 90% transmittance at 4000cm-1 when the sample is
present, to give peaks of the maximum deflection.
You should remember that this is an electromechanical instrument and
you should always make sure that you align the chart against the
calibration marks on the chart holder. In the more advanced instruments
an on-board computer stores a library of standard spectra, which can be
compared with your experimental spectrum.
2. F ourier transform IR (FT -IR) spectrometer: the value of IR
spectroscopy is greatly enhanced by Fourier transformation, named after
the mathematician J.B. Fourier. The FT is a procedure for inter-
converting frequency functions and time or distance functions. The IR
beam, composed of all the frequencies in the IR range, is passed through
the sample and generates interference patterns, which are then
transformed electronically into a normal IR spectrum. The advantages of
FT-IR are:
(a) rapid scanning speed - typically four scans can be made per minute,
allowing addition of the separate scans to enhance the signal-to-
noise ratio and improve the resolution of the spectrum;
(b) simplicity of operation - the reference is scanned first, stored and
then subtracted from the sample spectrum;
(c) enhanced sensitivity: the facility of spectrum addition from multiple
scans permits detection of smaller quantities of chemicals;

(d) the integral computer system enables the use of libraries of spectra
and simplifies spectrum manipulation, such as the subtraction of
contaminant or solvent spectra.
The procedures for running IR spectra on double-beam and FT

spectrometers are described in Box 28.1.
Sample handling
You can obtain IR spectra of solids, liquids and gases by use of the
appropriate sample cell (sample holder). The sample holder must be
completely transparent to IR radiation; consequently glass and plastic cells
cannot be used. The most common sample cells you will encounter are made
from sodium chloride or potassium bromide and you cannot use aqueous

solutions or very wet samples, otherwise the sample cells will dissolve. A
CD
typical range of sample cells is shown in Fig. 28.3 and for routine qualitative
work you will regularly use NaCI plates and KBr disks to obtain spectra of
solids and liquids. Solution cells and gas cells are utilized in more specialized
CD applications and require specific instructions and training.
Liquid samples
(a)
The most convenient way to obtain the IR spectrum of a pure, dry liquid
is to make a thin liquid film between two NaCl disks (plates). Since the
film thickness is unknown, this procedure is not applicable to quantitative
NaCI work.
window
Solid samples
If you were to place a fine powder between two NaCI plates, a usable
spectrum would not be obtained because the IR radiation would be scattered
by diffraction at the edges of the particles and would not pass through to the
detector. There are three solutions to this problem:
1. Mulls: in which the finely ground solid is mixed with a liquid, usually
Nujol® (liquid paraffin) or, less frequently, HCB (hexachloro-I,3-
butadiene). This mulling liquid does not dissolve the chemical but fills
NaCI the gaps round the edges of the crystals preventing diffraction and
windov
scattering of the IR radiation. Remember that these mulling liquids have
their own IR spectrum, which is relatively simple, and can be subtracted
either 'mentally' or by the computer. The choice of mulling liquid
depends upon the region of the IR spectrum of interest: Nujol® is a
(c) simple hydrocarbon containing only C-H and C-C bonds, whereas HCB
has no C-H bonds, but has C-Cl, C=C and C-C bonds. Examination of
Fig. 28.3 Cells for IR spectroscopy:
the separate spectra of your unknown compound in each of these
(a) demountable cell for liquid and solid films
and mulls; (b) solution cell; (c) gas cell. mulling agents enables the full spectrum to be analysed.
2. KBr disks: here the finely ground solid compound is mixed with
anhydrous KBr and squeezed under pressure. The KBr becomes fluid
and forms a disk containing the solid compound dispersed evenly within
it and suitable for obtaining a spectrum. The advantage of the KBr disk
technique is the absence of the spectrum from the mulling liquid, but the
disadvantages are the equipment required (Fig. 28.4) and the practice
required to obtain suitable transparent disks, which are very delicate and
rapidly absorb atmospheric moisture.
3. Thin solid films: here a dilute solution of the compound in a low-boiling-
point solvent such as dichloromethane or ether is allowed to evaporate
on a NaCI plate producing a thin transparent film. This method gives
excellent results but is slightly limited by solubility factors.
When you are recording spectra of mulls, KBr disks and thin solid films air is
used as the reference and they are suitable for qualitative analysis only. The
procedure for the preparation of liquid and solid films and mulls is described
in Box 28.2 and that for KBr disks in Box 28.3.
Interpretation of IR spectra
To identify compounds from their IR spectrum you should know at which
frequencies the stretching and bending vibrations occur. A detailed analysis
can be achieved using the correlation tables found in specialist textbooks. For
interpretation, the spectrum is divided into three regions.

Using spreadsheets
is very important and must be understood; it provides one of the most

common forms of error when copying formulae. Be sure to understand how
your spreadsheet performs these operations.
Naming blocks
When a group of cells (a block) is carrying out a particular function, it is
often easier to give the block a name which can then be used in all formulae
referring to that block. This powerful feature also allows the spreadsheet to
be more readable.
Graphics display
Most spreadsheets now offer a wide range of graphics facilities which are
easy to use and this represents an ideal way to examine your data sets rapidly
and comprehensively. The quality of the final graphics output (to a printer) is
variable but is usually sufficient for initial investigation of your data. Many
of the options are business graphics styles but there are usually histogram,
bar chart, X-Y plotting, line and area graphics options available. Note that
some spreadsheet graphics may not come up to the standards expected for
the formal presentation of scientific data (p. 343).
Printing spreadsheets
This is usually a straightforward menu-controlled procedure, made difficult
only by the fact that your spreadsheet may be too big to fit on one piece of
paper. Try to develop an area of the sheet which contains only the data that
you will be printing, i.e. perhaps a summary area. Remember that columns
can usually be hidden for printing purposes and you can control whether the
printout is in portrait or landscape mode, and for continuous paper or single
sheets (depending on printer capabilities). Use a screen preview option, if
available, to check your layout before printing. Most spreadsheets are now
WYSIWYG (What You See Is What You Get) so that the appearance on the
screen is a realistic impression of the printout. A 'print to fit' option is also
available in some programs, making the output fit the page dimensions.
Use as a database
Many spread sheets can be used as data bases, using rows and columns to
represent the fields and records (see Chapter 48). For many applications in
chemistry, the spreadsheet form of database is perfectly adequate and should
be seriously considered before using a full-feature database program.
310 Information technology and library resources

Practical Chemistry Techniques for Success

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Practical Skills in

We work with leading authors to develop the strongest educational materials

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First published 2002

© Pearson Education Limited 2002

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All rights reserved; no part of this publication may be

British Library Cataloguing-in-Publication Data

Typeset in Times Roman by 32

Fundamental laboratory techniques

The investigative approach

Analysis and presentation of data

Information technology and library resources

4.1 How to make up an aqueous solution of known concentration from a

27.5 Analysis of a sample: dilution factor 173

Practical skills form the cornerstone of chemistry. However, the diversity of

Metropolitan University), Jon Bookham, Susan Carlile, Jim Creighton, Sarah

We are grateful to the following for permISSIOn to reproduce copyright

Text extracts from the following databases: Analytical Abstracts, Chemical

Chapters 37-44 explain data analysis and presentation.

Chapters 45-49 cover information technology and library resources.

Chapters 50-56 deal with the vital subject of communicating information.

Ar relative atomic mass

All knowledge and theory in science has originated from practical

~ You will get the most out of laboratory work if

The main points to remember are:

Fundamental laboratory techniques 3

All experimental observations and data should be recorded in a notebook

Presenting more advanced practical work

4 Fundamental laboratory techniques

To prepare overhead transparencies for oral presentations, you can use

Fundamental laboratory techniques 5

Health and safety law requires institutions to provide a working environ-

~ All practical work must be carried out with safety in

6 Fundamental laboratory techniques

LABORATORY RISK ASSESSMENT

This sheet must be completed before commencing each experiment

REAGENT AND HAZARD (H) and RISK (R) CODES

A = Corrosive/irritant M = Microbiological hazard 1 =low/nohazardJrisk

2. Precautions: containment and protection

Stage of Experiment Containment Personal Additional Information

Fig.2.2 An example of a laboratory hazard assessment form.

Basic rules for laboratory work

Fundamental laboratory techniques 7

Fig.2.3 Warning labels for specific chemical

8 Fundamental laboratory techniques

Measuring and dispensing liquids

Table 3. 1 Criteria for choosing a method for measuring out a liquid

Pasteur pipette 1-5 mL Low Convenient

*If calibrated correctly and used properly (see p. 10).

Conical flasks, beakers, measuring cylinders and volumetric flasks

Fundamental laboratory techniques 9

Conical flasks and beakers

Measuring cylinders and volumetric flasks

~ For safety reasons, it is no longer permissible to

10 Fundamental laboratory techniques

Air displacement and positive displacement pipettors may be:

Fig. 3.4 A pipettor - the Gilson Pipetman'P.