Chromatographic Methods

392 Pages·1974·10.794 MB·English

by  R. Stock, C. B. F. Rice (auth.)

CHROMATOGRAPHIC METHODS
Chromatographic
Methods
R. Stock
Head of Department of Physical Sciences
Trent Polytechnic, Nottingham
C. B. F. Rice
Principal Lecturer in Chemistry
Liverpool Polytechnic
SPRINGER-SCIENCE+BUSINESS MEDIA. B.Y.
© 1963,1967,1974 R. Stock and C. B. F. Rice
Originally published by Chapman and Hall in 1974
Softcover reprint of the hardcover 1st edition 1974
Typeset by Santype Ltd (Coldtype Division)
Salisbury, Wiltshire
ISBN 978-0-412-20810-2 ISBN 978-1-4899-3360-7 (eBook)
DOI 10.1007/978-1-4899-3360-7
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Contents
Preface to First Edition page vi
Preface to Third Edition vii
1 Introduction 1
2 Liquid-phase Chromatography on Columns 18
3 Paper Chromatography; Zone Electrophoresis 106
4 Gas Chromatography 177
5 Thin-layer Chromatography 279
6 Model Experiments in Chromatographic Techniques 318
Appendix 360
Bibliography 371
Index 376
Preface to First Edition
The various methods of separating mixtures which are
grouped under the general name 'chromatography' are now
well known and widely used. Since the inception of
chromatography as a column technique in 1903, the principal
landmarks in its progress have been its virtual rediscovery in
1930, the invention of synthetic ion-exchange resins in 1935,
the introduction of paper chromatography in the early
1940's, and fmally the development of gas-solid and gas
liquid chromatography in the late 1940's and early 1950's.
Subsequent expansion in the use of chromatographic
methods has been rapid and continuous, with the result that
in the last fifteen years a substantial volume of literature on
the subject has appeared, dealing not only with particular
separations but also in much specific detail with improve
ments in technique.
Many specialist books have been published. Some are
concerned only with particular aspects of the subject. Others
are essentially literature surveys which are usually very
comprehensive (though somewhat uncritical), and hence
rather formidable to someone seeking an introduction to
chromatography.
The present book aims to present a short account of
techniques in current use. Emphasis has been given to paper
and gas chromatography because they are now the most
widely used methods, aithough thin-layer chromatography
has recently become extremely popular. The treatment is
dictated by the need to stress practical considerations, and is
designed to show how the various methods complement
one another. Details are given of some model experiments
vi
Preface to First Edition . vii
based on the experience of the authors in teaching the
subject. The inclusion of student experiments in gas chro
matography is regarded as an important development. Zone
electrophoresis, though not strictly a chromatographic tech
nique, is nearly enough related to be included. An attempt
has been made to give some guidance as to the choice of
method for a particular purpose, and to assess the relative
values of the different procedures.
The authors gratefully acknowledge permission to repro
duce published information, diagrams, or photographs from:
B.D.H. Ltd., The Dow Corporation, Evans Electroselenium
Ltd., Mr. G. F. Harrison of Associated Octel Co. Ltd., Dr.
C. S. Knight of W. R. Balston & Co. Ltd., Dr. S. W. S.
McCreadie and Dr. A. F. Williams of Nobel Division, I.C.1.
Ltd., The Permutit Co. Ltd., H. Reeve Angel & Co. Ltd., the
Journal of Applied Chemistry, the Journal of Chemical
Education, and Nature. Thanks are also due to Dr. S. J. Gregg
for much helpful criticism and discussion, to Dr. G. Skirrow
for suggesting certain of the gas chromatography experiments,
and, finally, to Mrs. R. Wiggins for preparation of the
typescript.
College of Technology R. STOCK
Liverpool C. B. F. RICE
PREFACE TO THIRD EDITION
An evaluation of the literature relating to chromatography by
Janak (Janak, J., J. Chromatog., 1973,78, 117) has shown
that since 1967 the number of research papers on paper and
thin-layer chromatography has fallen, the number on gas
chromatography, after an initial fall, has started to rise again,
and the number on liquid chromatography has shown a
spectacular increase. The number of research papers pub-
viii • Preface to Third Edition
lished in a particular period is not necessarily an exact
measure of the extent to which a method is actually used
during the time in question, but it is probably indicative of
trends.
The increase in interest in liquid, chromatography on
columns, although due in part to the continued development
of gel chromatography, is due mainly to the application of
the methods of gas chromatography in what is now called
high efficiency liquid chromatography, accompanied by
increased sophistication in instrumentation and improve
ments in manufacture of chromatographic media.
These trends are reflected in the third edition of this book.
The sections on gas chromatography and gel chromatography
have been rewritten and expanded, and a section on high
efficiency liquid chromatography has been included, but the
remaining section, apart from minor revisions and correc
tions, are substantially as in the second edition. Throughout
the book SI units are now used and references to commercial
products have been brought up to date. The bibliography has
been made more selective and the list of visual aids for
teaching has been extended.
February 1974 R. STOCK
Trent Polytechnic, Nottingham
C. B. F. RIC E
Liverpool Polytechnic
Chapter One
Introduction
Classification of chromatographic methods
Chromatography is the name given to a particular family of
separation techniques of great effectiveness. The original
method was described in 1903 by Tswett, who used it for the
separation of coloured substances, and the name chromatog
raphy stems from this. However, the limitation to coloured
compounds never really obtained, and most chromatographic
separations are nowadays performed on mixtures of
colourless substances, including gases.
Like fractional distillation, chromatography relies on the
relative movement of two phases, but in chromatography one
is fixed and is known as the stationary phase; the other is
known as the mobile phase. Chromatographic methods may
be classified first according to the nature of the mobile phase
and, second, according to the nature of the stationary phase.
The mobile phase may be a liquid or a gas, and the stationary
phase may be a solid or a liquid. There are thus four main
sub-divisions of the chromatographic process, as set out in
Table 1.1. The system is called adsorption chromatography if
the stationary phase is a solid, and partition chromatography
if it is a liquid.
There are several variants of the four main types, as shown
below:
1. Liquid-solid
'Classical' adsorption chromatography
Thin-layer chromatography
Ion-exchange chromatography
Gel chromatography
I\.) 9 a ~ 8' ~ i c)' ~ s-o ~
e
il y
mob d aph
e ~ eous hase -liquiatogr
as aspasm
h G Go
N y p 4) Chr
t TITIOionar (
PARuid stat obile uid aphy
ods ~~ Liq ~ quid mphase uid -liqmatogr
Meth ~; ) Li Liqhro
1.1 of RG P C
Table sification HROMA;'-O I mobile d aphy
Clas I C e ~ seous phase as-solimatogr
as Ga Go
N ph ) hr
TIOry (2 C
~ ADSORPSolid stationa r-- Liquid mobile ) phase Liquid-solid hromatography
1 C
(