Buffers for pH and

Metal Ion Control

Buffers for pH and
Metal Ion Control

D. D. Perrin
John Curtin School of Medical Research
Australian National University
Canberra

Boyd Dempsey
Faculty of Military Studies
University of New South Wales
Royal Military College
Duntroon

London
CHAPMAN AND HALL
First published in 1974
by Chapman and Hall Ltd
11 New Fetter Lane, London EC4P 4EE
First issued as a Science Paperback 1979
© 1974 D.D. Perrin and Boyd Dempsey
Typeset by Santype Ltd (Coldtype Division)
Salisbury, Wiltshire

ISBN-J3: 978-0-412-21890-3 e-ISBN-13: 978-94-009-5874-6

DOl: /0./007/978-94-009-5874-6

This paperback edition is

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without the publisher's prior consent in any form of
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on the subsequent purchaser
All rights reserved. No part of
this book may be reprinted, or reproduced
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from the publisher.

Distributed in the US.A.

by Halsted Press, a Division
of John Wiley & Sons, Inc, New York
Contents

Preface page viii

1. Introduction 1
1.1 The concept of buffer action 1
1.2 Why are buffers needed? 2
1.3 Some naturally occurring buffers 3
2. The Theory of Buffer Action 4
2.1 Equilibrium aspects 4
2.2 Activity effects 6
2.3 Effect of dilution 7
2.4 Salt effects 8
2.5 Ampholytes and zwitterions 10
2.6 Buffer capacity 10
2.6.1 Buffer capacity of a polybasic acid 12
2.7 Pseudo buffers 15
2.8 Self buffers 15
2.9 Mixtures of buffers 17
2.10 Temperature dependence 18
2.11 Effect of pressure on buffers 18
2.12 Further reading 19
3. Applications of pH Buffers 24
3.1 Factors governing the choice of a buffer 24
3.2 Measurement of pH 25
3.3 Biochemistry and biology 27
3.4 Spectroscopy 32
3.5 Buffers for special applications 33
3.5.1 Volatile buffers 33
3.5.2 Buffers for electrophoresis 34
3.5.3 Buffers for complexometric titrations 34
3.5.4 Buffers for chromatography 35
vi . Contents

3.5.5 Buffers for polarography 35

3.5.6 Buffers for proton magnetic resonance studies 36
3.5.7 Buffers for solvent extraction 36
3.5.8 Isotonic phannaceutical buffers 37
3.5.9 Miscellaneous 38
4. Practical Limitations in the Use of Buffers 55
4.1 Chemical problems 55
4.2 Biological effects 58
4.3 Influence on chemical reactions 60
5. New pH-Buffer Tables and Systems 62
5.1 On calculating buffer composition tables 62
5.1.1 Buffers of constant ionic strength.
No added electrolyte 62
5.1.2 Constant ionic strength buffers with added electrolyte 64
5.1.2.1 Preparation of amine buffers of constant ionic
strength 65
5.1.3 Buffers by direct titration of weak bases or acids with
strong acids or bases 65
5.2 On designing a new pH-buffer system 69
6. Buffers for use in Partially Aqueous and
Non-Aqueous Solvents and Heavy Water 77
6.1 pH* Scales 78
6.2 pH* Buffers 78
6.3 The measurement of pH* 79
6.4 A universal pH scale 80
6.5 The pD scale and the measurement of pD 81
6.6 The use of pH* and pD buffers 82
6.6.1 The determination of dissociation constants of acids 82
6.6.2 Rate studies in heavy water 82
6.7 Surfactants 83
7. Metal-ion Buffers 94
7.1 The concept of pM 94
7.2 Uses of metal-ion buffers 95
7.3 Calculation of pM 96
7.4 pH-Independent metal-ion buffers 99
7.5 Effects of pH buffer substances on pM 101
7.6 Anion buffers 102
7.7 Redox buffering 103
8. Purification of Substances Used in Buffers 109
9. Preparation of Buffer Solutions 117
 Contents· vii

10. Appendices 123

Appendix I. Tables for constructing buffer tables 123
Appendix II. Composition-pH tables of some commonly
used buffers 128
Appendix III. Thermodynamic acid dissociation
constants of prospective buffer substances 157
Appendix IV. The Henderson-Hasselbalch equation 164
References 167
Index 173
Preface

This book is intended as a practical manual for chemists,

biologists and others whose work requires the use of pH or
metal-ion buffers. Much information on buffers is scattered
throughout the literature and it has been our endeavour to
select data and instructions likely to be helpful in the choice
of suitable buffer substances and for the preparation of
appropriate solutions. For details of pH measurement and the
preparation of standard acid and alkali solutions the reader is
referred to a companion volume, A. Albert and E. P.
Serjeant's The Determination of Ionization Constants (1971).
Although the aims of the book are essentially practical, it
also deals in some detail with those theoretical aspects
considered most helpful to an understanding of buffer
applications. We have cast our net widely to include pH
buffers for particular purposes and for measurements in
non-aqueous and mixed solvent systems. In recent years there
has been a significant expansion in the range of available
buffers, particularly for biological studies, largely in conse-
quence of the development of many zwiUerionic buffers by
Good et al. (1966). These are described in Chapter 3.
However, there are very many substances that could be, or
have been, of use as buffers, and Appendix III lists some of
these. Chapter 5 shows how new pH-buffer tables can be
constructed from the thermodynamic pKa values, and some
simple computer programmes are included to facilitate the
necessary calculations. Tables and worked examples are given
for use if a computer is not available.
In view of the importance of metal ion concentrations,
particularly in biological work, we have considered it
appropriate to include a section on metal-ion buffers. These
buffers may also be useful in preparing convenient standards
for ion-selective electrodes.

Canberra, D. D. Perrin
August, 1973 Boyd Dempsey