TLC - Vol 1a - Jork, Funk, Fisher & Wimmer.pdf
(
17216 KB
)
Pobierz
<!DOCTYPE html PUBLIC "-//W3C//DTD HTML 4.01//EN" "http://www.w3.org/TR/html4/strict.dtd">
'
Waiter Fischer, Hans Wimmer
Walter Fischer, Hans Wimmer
Thin-Layer
Chromatography
Reagents and Detection Methods
Volumel
Volume2
Radiometric Detection Methods
(in preparation)
Volume
la
Biochemical and Biological Detection
(in preparation)
Translated by Frank and Jennifer A. Hampson
D1stnbutlon
VCH Verlagsgesellschaft, P 0 Box 101161, D 6940 Wemhenn (Federal Repubhc of Gennany)
Switzerland VCH Verlags-AG, P 0 Box, CH-4()2(] Base] (Swttzcrland)
Umted Kmgdom and Ireland VCH Publishers (UK) Ltd , 8 Welhngton Court,
Welhngton Street, Cambndge CBllHW (England)
USA and Canada VCH Pubhsben., Smte 909,220 East 23rd Street, New York.
NY 10010-4606 (USA)
.
ISBN 3-527-27834-6 (VCH, Wemhe!m)
ISBN 0-89573-876-7 (VCH, New York)
J<orewora
Prof W Funk
Prof Dr H
Jorlr,;
Fachbere:tch Teclumches Gesundbettswesen
U
mversttit des Saarlandes
der Fachhochschule Gte8en-Fnedberg
Fachberetch 14
WtesenstraBe 14
Stadtwa1d
D-6300 G1e6en
D-6600 Saarbriick.en
Thin-layer chromatography as practiced today seems to exist in two forms. Some
Hans Wnumer
Dr W FISCher
Eckhardt-StraBe 23
sctentJ.sts consider TLC to be a qualitative separation tool for simple mixtures
cfo E Merck
D-6100 Dannstadt
Abteilung V
Reag
SPA
where speed, low cost,
an~ t~plicity
are its
"~ues.
Others regard TLC as a
Frankfurter StraBe 250
,.,,;, ·
1
'' · · · •
m;Ytm~
woth "
high sample throughput because ofparallel sample processing, and as a technique
that can tolerate cruder samples than column methods because the stationary
Thts book
was
carefully produced Nevertheless, authors, translator and pubhsher do not warrant
phase is disposable, and which provides flexibility
in
the method and chmce of
the mfonnauon oontawed therein to be free of errors Readers are adVIsed to keep 10 mmd that
statements, data, tllustrattons, procedural detatls or other 1tems may madvcrtently be maccurate
detection since at the time of detection the separation is static and the layer open
to mspectJ.on. Both groups of scientists use the same approaches and employ the
:,
.'o, '
on\v
lh
>0
·'
··'"
Product!on Manager Dtpl -Ing (FH) Hans JOrg Mater
so m an optimized way. There remains an infonnatJ.on gap winch good books can
llJJ to re-educate the sctentthc communrry orme currem stanuing 01
t
LL. 1 am
Library of Congress Card No
deltghted to affinn that the present book takes a needed step in tlus direction. As
89-16558
more scientists become acquainted with the modern practice ofTLC they wdl need
reliable and unbiased sources ofinfonnabon on the mynad offactors that influence
Bntlllh L1brary Catalogwng-m-Pubhcabon Data
Thiolayer dlromalograplly reagents and detectiOn methods
quantitahon of TLC chromatograms to avotd common pitfalls that follow in
Vol la phyStcal and cheiD!cal detectton methods
1 Thm layer chromatography
the wake of any technologtcal advance. From phystcal pnnctples, to working
Instruments, to tne mernooo ogica1
-; ·
~
pluluU.J•
u
~ u~
au
~4J3~~i9~:umut
will
fin..t
"""h infnnnation and iust as
im
ortantlv. exoerience dtsblled
,...J,
""
roe-
mto tlus book.
Deutsche Btbhothek Catalogwng-m-Pubhcatwn Data
It
seems to be a fact of hfe that real samples are too dilute or too complex for
lld!l-Lityer cbfOIDatogrliphy reagents and detectton methods I
dtrect analyses no matter what new technology IS avatlable simply because the
Hellmut Jork
- Wemhe1m , Basel (SWitzerland) , Cambndge
demand for analyttcalmfonnation is being contmuously ratsed to a lugher level.
, New York, NY VCH
NE Jork, HeUmut [Mttverf]
At this mterface chemical mtuition has always played an important role. SelectJ.ve
chetn1cal reactions nrov1de the methods to marunulate a sample to reveal the
·"
mformation desued. They provide the means to increase the response of an
~<.:~~n~_am~ts, rear~~nh
·~
"'
ISBN 0-89573-876-7 (New York
)
.
targetmg certam components of the sample to respond to a selected detector.
©
VCH Verlagsge5ellschaft mbH, D-6940 Wemheun (Federal Repubhc of Germany), 1990
Mtcropreparative chemistry and TLC have a long htstory of association because
of the convenience of these reactions when performed with a static sample and
Pnnted on llCld-free paper
because m TLC the separation and detection processes can be treated as separate
All
nghts
reserved (mcludmg those of translation mto other languages) No
part
of thts book may be
reproduced many fonn - by photoprmt, mlcromm. or any other means - nor transrrutted or translated
steps and optliJllZed independently of each other. A further important character
~~J. ...;tt~·,;,:.:..l!lSI.on
from the nubhshers Rel!lstered names, trademarks,
etc used m this book. even
when
not spectfica11y marked
as
such, are not to be ronsuiered unproter.ted
by law
together provtding an mtegrated source to the most Important chemical reactions
Composltlon and Pnntang Wtesbadener Gr.tplnscbe Betnebe GmbH. D-6200 Wtesbaden
ava1lable and the details of therr apphcatlon to particular sample types. Unttl new
Bookbmdmg OeorgKrankl, 0-6148 Heppenhenn
detectwn pnnC!ples are avatlable, these reactions represent the most practical and,
Prmted mthe Federal Republic of Germany
VI
Foreword
Pretace
in many cases, an elegant solution to difficult analytical problems. This book
should serve to revive interest
in
this area and to provide a methodological source
for their practice.
The power of TLC is in its flexibility as a problem solving tool.
As
the problems
in analysis become more complicated the sophistication by which we approach
This book is the result of cooperation between four colleagues, who have been
working in the field
ofthin~layer
chromatography for many years and, in particu-
those problems is ever increasing. However,
it
behooves us as analytical chemists
Jar, took an active part in
the
development from hand-coated TLC plates to
·
not to forget our fundamental training in chemistry and to apply those principles
nlo>;
"~
·
.chmma-
.i!ohl"
·;;,;._
0 ,.
.
~-,
tography. This development was accompanied
by
improvements in the field of
this
ok.
detection of
the
separated zones. In particular, it became necessary to
be
able
C.
F. Poole
to deal with ever decreasing quantities of substance, so that the compilation
Department of Chemistry
"Anf8.rbereagenzien" by E. Merck, that had been available as a brochure for
Wayne State University
many, many years, no longer represented the state of the art of thin-layer
Detroit, MI 48202
It
was against this background and in view of the fact that there is at present
no contemporary monograph on mm-layer chromatograPfiYllUilmis uuul\. w<ts
produced.
It
is intended as an introduction to the method, a reference book, and
a laboratory handbook
in
one, i.e., far more than just a "Reagent Book".
The first part of the book consists of a detailed treatment of the fundamentals
of thin-layer chromatography, and of measurement techniques and apparatus for
the qualitative and quantitative evaluation of thin-layer chromatograms. ln situ
prechromaiOgrapmc oenvauza ion tecnniques useu
w
i~prove
""' · ·
· , t1 ·
~rea!U':
the
!U':O~it'vitv
ofdetection and to enhance the precision
of the subsequent quantitative analysis are summarized in numerous tables.
Particular attention has been devoted to the fluorescence methods, which are
now of such topicality, and to methods of increasing and stabilizing the fluores-
cence emissions. Nowhere else in the literature is there so much detailed infor-
mation to
be
found as in the first part ofthis book, whose more than 600 literature
rpfp,..,.n.-...
mav serve to stimulate the reader to enlarl!e his or her own knowledge.
Nor has a general introduction to the microchemical postchromatographic
This
s:~nd
part
wi~h
its 80 worked-through and checked
d~tection
methods
forms the foundation of a collection of reagent reports (monographs), which will
be
extended to several volumes and which is also sure to
be
welcomed by workers
who carry out derivatizations in the fields of electrophoresis and high-pressure
l~quid chromatograph~. A.l~ngside
details of the
rea~nts r~ui~~ an~~heir ha~d-
"•
.•..
concerned.
Wherever possible, dipping reagents have been employed instead of the spray
reagents that were formerly commonplace. These make it easier to avoid contami-
IX
Preface
Vlll
Preface
In spite of all our care and efforts we are bound to have made mistakes. For
nating the laboratory, because the coating of the chromatogram
with
the reagent
this reason we would like to ask TLC specialists to communicate to us any errors
takes place with less environmental pollution and lower health risks; furthermore,
and any suggestions they may have for improving later volumes.
it is more homogeneous, which results in higher precision in quantitative ami.lyses.
It
is possible that the solvents suggested will not be compatible with all the
Saarbriicken, Giessen and Darmstadt,
Hellmut Jork
substances detectable with a particular reagent, for instance, because the chro-
malographically separated substances or their reaction products are too soluble.
Werner Funk
Waiter Fischer
October 1989
Therefore,
it
should be checked in each case whether
it
is possible to employ the
H'
,w;
conutuons suggesu;u WlluOUl ffiOUillCauon. vve illiVe uone
llllS
m eacn report
wr
one chosen class of substance by working through an example for ourselves and
have documented the results in the «Procedure Tested"; this includes not only the
exact chromatographic conditions but also details concerning quantitation and
the detection limits actually found. Other observations are included as «Notes".
Various types of adsorbent have been included in these investigations and their
• • • 1
i~ a1~~
n;:pUJ.u;u. u an
au~onx::m
is DOL memioneu n orny means mat
we did not check the annlication of the rea ent to that tvne
f 1
t
that
1.-i
the reagent cannot be employed on that layer.
Since, in general, the reagent report includes at least one reference covering each
substance or class of substances,
it
is
possible to use Part
ll
of this book with its
ea. 750 references as a source for TLC applications. Only rarely are earlier refer-
ences (prior to
1960),
which were ofimportance for the development ofthe reagent,
. •rlh.
There is no need to emphasize that many helpful hands are required in the
compilation of such a review. Our particular thanks are due to Mrs. E. Kany, Mrs.
I.
Klein and Mrs. S. Netz together with Dipl.-Ing. M. Heiligenthal for their
conscientious execution of the practical work.
We would also like to thank the graduate and postgraduate students who helped
to check the derivatization reactions and Mrs.
U.
Enderlein, Mrs. E. Otto, and
Mrs. H. Roth, whose capable hands took care of the technical preparations for
the book and the production of the manuscript. We would particularly like to
.
.
~'
of the reaction paths for the reagent reports. Our thanks are also due to Dr. F.
Hampson and Mrs.
J.
A.
Hampson for translating the German edition ofthe book
into English.
We thank the Baron,
J. T.
Baker, Camag, Desaga, Macherey-Nagel and E.
Merck companies for their generous support of the experimental work.
Verlagsgesellschaft for the realization of our concepts and for the design and
presentation of the book and for the fact that this work has appeared in such a
short time.
'
Contents
Part I
Methods of Detection
Introduction
I
3
2
Physical Methods of Detection
9
2.1
General
9
~
2.2.1
Visual Detection
2.2.2
Fluorescence and Phosphorescence Indicators .
10
2.2.3
Photometric Measurement of Absorption .
17
Apparatus
2.2.3.1
17
2.2.3.2
Principles of Measurement.
30
2.2.3.3
Quantitative Analysis
34
2.3
The Detection of Fluorescent Substances
37
2.3.1
General
37
J
2.3.3
Fluorimetric Determinations .
38
2.4
Detection of Radioactively Labelled Substances
40
2.5
Nondestructive Detection Using Other Physical Methods.
42
2.5.1
Spectral Phenomena
42
2.5.2
Wetting and Solubility Phenomena .
42
'"
2.5.4
Treatment with Iodine.
46
3
Chemical Methods of Detection .
55
3.1
In situ Prechromatographic Derivatization
56
3.1.1
Oxidation and Reduction
58
3.1.2
Hydrolysis
62
3.1.3
Halogenation .
64
1' 1tratton ana utazotlzatwn ..
66
·'-'·'
3.1.5
Esterification and Etherification
68
3.1.6
Hydrazone Formation.
71
3.1.7
Dansylation
72
Plik z chomika:
Tkanczak
Inne pliki z tego folderu:
Vol 2 2000.pdf
(116740 KB)
Structure elucidation by NMR 3ed - Breitmaier.pdf
(12052 KB)
Modern derivatization methods for separation sciences 1999 - Toyo'oka.pdf
(6446 KB)
Vol 3 2000.pdf
(105832 KB)
HPLC and CE -Principles and Practice 1997 - Weston & Brown.pdf
(14893 KB)
Inne foldery tego chomika:
Dawkins
Death Note
eBooks - Kryminalistyka i medycyna sądowa
Ergo Proxy
Filmy Korea Południowa
Zgłoś jeśli
naruszono regulamin