Poole Instrumental Thin-Layer Chromatography

1. Auflage 2014
ISBN: 978-0-12-417284-5
Verlag: Elsevier Science & Techn.
Format: EPUB
Kopierschutz: 6 - ePub Watermark

E-Book, Englisch, 652 Seiten

Reihe: Handbooks in Separation Science

Instrumental Thin-Layer Chromatography delivers comprehensive coverage of this separation tool with particular emphasis on how this tool can be used in advanced laboratories and integrated into problem-solving scenarios. Significant improvements in instrumentation have outpaced the development of information resources that describe the latest state-of-the-art and demonstrate the full capabilities of TLC. This book provides a contemporary picture of the fundamentals and practical applications of TLC at a level suitable for the needs of professional scientists with interests in project management where TLC is a common tool. Compact, highly focused chapters convey essential information that defines modern TLC and how it can be effectively implemented in most areas of laboratory science. Numerous figures and tables provide access to material not normally found in a single source yet are required by working scientists. - Contributions written by recognized authoritative and visionary experts - Focuses on state-of-the-art instrumental thin-layer chromatography and advanced applications across many areas - Provides guidance on the analysis of complex, dirty mixtures of compounds - Offers a cost-effective analytic technique for laboratories working under strict budgets

Professor Colin Poole is internationally known in the field of thin-layer chromatography and is an editor of the Journal of Chromatography and former editor of the Journal of Planar Chromatography - Modern TLC. He has authored several books on chromatography, recent examples being The Essence of Chromatography published by Elsevier (2003), and Gas Chromatography published by Elsevier (2012). He is the author of approximately 400 research articles, many of which deal with thin-layer chromatography, and is co-chair of the biennial 'International Symposium on High-Performance Thin-Layer Chromatography”.

Autoren/Hrsg.

Poole, Colin

Inhaltsverzeichnis

1;Front Cover;1
2;INSTRUMENTAL THIN-LAYER CHROMATOGRAPHY;4
3;Copyright;5
4;Contents;6
5;Contributors;8
6;Chapter 1 - Milestones, Core Concepts, and Contrasts;12
6.1;1.1 INTRODUCTION;12
6.2;1.2 MILESTONES;13
6.3;1.3 ATTRIBUTES OF A PLANAR FORMAT;15
6.4;1.4 CONSEQUENCES OF CAPILLARY-CONTROLLED FLOW;18
6.5;1.5 SOLVENT-STRENGTH GRADIENTS;30
6.6;1.6 MULTIDIMENSIONAL SEPARATIONS;33
6.7;1.7 CONCLUSIONS;36
6.8;References;37
7;Chapter 2 - High-Performance Precoated Stationary Phases;42
7.1;2.1 INTRODUCTION;42
7.2;2.2 INORGANIC OXIDE SORBENTS;43
7.3;2.3 CHEMICALLY BONDED SORBENTS;50
7.4;2.4 CELLULOSE;56
7.5;2.5 CHIRAL SORBENTS;57
7.6;2.6 CONCLUSIONS;59
7.7;References;60
8;Chapter 3 - Ultrathin and Nanostructured Stationary Phases;64
8.1;3.1 INTRODUCTION;64
8.2;3.2 MONOLITHIC SILICA GELS;67
8.3;3.3 MONOLITHIC POLYMERS;69
8.4;3.4 ELECTROSPUN NANOFIBERS;70
8.5;3.5 NANOSTRUCTURED THIN FILMS;72
8.6;3.6 CARBON NANOTUBE-TEMPLATED LAYERS;75
8.7;3.7 ADVANCED INSTRUMENTATION AND TECHNIQUES;77
8.8;3.8 CONCLUSIONS;79
8.9;References;80
9;Chapter 4 - Automated Multiple Development;84
9.1;4.1 INTRODUCTION;84
9.2;4.2 APPLICATIONS;94
9.3;4.3 FUTURE TRENDS;105
9.4;4.4 CONCLUSIONS;108
9.5;References;109
10;Chapter 5 - Forced-Flow Development in Overpressured Layer Chromatography;118
10.1;5.1 INTRODUCTION;118
10.2;5.2 ANALYTICAL AND PREPARATIVE OPLC PROCESSES;119
10.3;5.3 INSTRUMENTS AND LAYERS FOR OPLC;123
10.4;5.4 MAIN CHARACTERISTICS OF OPLC;127
10.5;5.5 ANALYTICAL AND PREPARATIVE APPLICATIONS;133
10.6;5.6 CONCLUSION;140
10.7;References;140
11;Chapter 6 - Pressurized Planar Electrochromatography;146
11.1;6.1 INTRODUCTION;146
11.2;6.2 THEORETICAL BACKGROUND;147
11.3;6.3 DEVELOPMENT OF EQUIPMENT FOR PPEC;151
11.4;6.4 ADVANTAGES OF PPEC;159
11.5;6.5 APPLICATIONS;168
11.6;6.6 CHALLENGES AND CONCLUSIONS;174
11.7;References;174
12;Chapter 7 - Theory and Instrumentation for In situ Detection;178
12.1;7.1 INTRODUCTION;178
12.2;7.2 THEORY FOR IN SITU DENSITOMETRIC DETECTION;179
12.3;7.3 INSTRUMENTATION FOR IN SITU DENSITOMETRIC DETECTION;187
12.4;7.4 IN SITU MASS-SPECTROMETRY;197
12.5;7.5 IN SITU RADIOISOTOPE DETECTION;198
12.6;References;199
13;Chapter 8 - Staining and Derivatization Techniques for Visualization in Planar Chromatography;202
13.1;8.1 PROBLEM OVERVIEW;202
13.2;8.2 REAGENT APPLICATION, EQUIPMENT, AND PROTOCOLS;204
13.3;8.3 TECHNIQUES FOR HEATING OR DRYING LAYERS AFTER DEVELOPMENT;212
13.4;8.4 COMMON DETECTION PROTOCOLS FOR TARGET COMPOUNDS;215
13.5;8.5 CONCLUSIONS;246
13.6;References;246
14;Chapter 9 - Advanced Spectroscopic Detectors for Identification and Quantification: UV–Visible, Fluorescence, and Infrared ...;250
14.1;9.1 INTRODUCTION;250
14.2;9.2 ADVANCED SPECTROSCOPIC DETECTORS FOR IDENTIFICATION AND QUANTIFICATION;251
14.3;9.3 CONCLUSION;258
14.4;References;258
15;Chapter 10 - Advanced Spectroscopic Detectors for Identification and Quantification: Mass Spectrometry;260
15.1;10.1 INTRODUCTION;260
15.2;10.2 CLASSIFICATION OF TLC-MS TECHNIQUES;261
15.3;10.3 INDIRECT SAMPLING TLC-MS;261
15.4;10.4 DIRECT SAMPLING TLC-MS;265
15.5;10.5 HIGH-THROUGHPUT TLC-MS DEVICES AND QUANTIFICATION ANALYSIS;279
15.6;10.6 CONCLUSION;281
15.7;References;282
16;Chapter 11 - Effects-Directed Biological Detection: Bioautography;290
16.1;11.1 INTRODUCTION;290
16.2;11.2 APPLICATIONS OF BIOAUTOGRAPHIC TESTS;295
16.3;11.3 PERSPECTIVES;314
16.4;References;314
17;Chapter 12 - Solvent Selection and Method Development;324
17.1;12.1 INTRODUCTION;324
17.2;12.2 PROBLEM DEFINITION;325
17.3;12.3 MODE SELECTION;330
17.4;12.4 MOBILE PHASE SELECTION;334
17.5;12.5 CONCLUSIONS;359
17.6;References;359
18;Chapter 13 - Validation of Thin Layer Chromatographic Methods;362
18.1;13.1 INTRODUCTION;362
18.2;13.2 METHOD VALIDATION USING THE CLASSIC APPROACH;363
18.3;13.3 ALTERNATIVE METHOD VALIDATION APPROACH USING ACCURACY PROFILES;372
18.4;13.4 CONCLUSION;382
18.5;References;382
19;Chapter 14 - Separation of (Phospho)Lipids by Thin-Layer Chromatography;386
19.1;14.1 INTRODUCTION;386
19.2;14.2 SEPARATION OF LIPIDS BY TLC;389
19.3;14.3 APPLICATIONS;393
19.4;14.4 MALDI FOR MS DETECTION;406
19.5;14.5 SUMMARY AND OUTLOOK;410
19.6;Acknowledgments;410
19.7;References;411
20;Chapter 15 - Applications in Food Analysis;418
20.1;15.1 INTRODUCTION;418
20.2;15.2 HPTLC IN FOOD ANALYSIS;420
20.3;15.3 SAMPLE PREPARATION, HYPHENATION, AND NEW POSSIBILITIES;424
20.4;15.4 SUITABILITY AND CAPABILITIES;432
20.5;15.5 CONCLUSION;435
20.6;References;435
21;Chapter 16 - Environmental Applications;442
21.1;16.1 INTRODUCTION;442
21.2;16.2 HPTLC IN ENVIRONMENTAL ANALYSIS;443
21.3;16.3 CONCLUSION;455
21.4;References;456
22;Chapter 17 - Pharmaceutical Applications of High Performance Thin Layer Chromatography;462
22.1;17.1 INTRODUCTION;462
22.2;17.2 QUANTITATIVE ANALYSIS;464
22.3;17.3 PHARMACEUTICAL APPLICATIONS;464
22.4;17.4 CONCLUSIONS;488
22.5;References;488
23;Chapter 18 - Utility of Thin-Layer Chromatography in the Assessment of the Quality of Botanicals;490
23.1;18.1 INTRODUCTION;490
23.2;18.2 METHOD VALIDATION;494
23.3;18.3 HYPHENATED TECHNIQUES AND CHEMOMETRICS IN HERBAL ANALYSIS;497
23.4;18.4 APPLICATIONS OF TLC IN THE FIELD OF HERBAL PRODUCTS/BOTANICALS;499
23.5;18.5 USE OF HPTLC BY THE AMERICAN HERBAL PHARMACOPOEIA;511
23.6;18.6 RECENT ADVANCES IN HPTLC-BASED QUALITY CONTROL OF BOTANICALS AND DIETARY SUPPLEMENTS;511
23.7;18.7 CONCLUSIONS;512
23.8;References;512
24;Chapter 19 - Analysis of Plant Material;516
24.1;19.1 INTRODUCTION;516
24.2;19.2 SINGLE DEVELOPMENT;517
24.3;19.3 SPECIAL DEVELOPMENT TECHNIQUES;525
24.4;19.4 COUPLING PLANAR CHROMATOGRAPHY WITH COLUMN CHROMATOGRAPHY;535
24.5;19.5 FORCED-FLOW DEVELOPMENT;535
24.6;19.6 CHEMICAL FINGERPRINTING OF PLANT MATERIALS;537
24.7;19.7 EFFECT-DIRECTED ANALYSIS OF PLANT MATERIALS;541
24.8;19.8 IMAGE PROCESSING;549
24.9;References;551
25;Chapter 20 - Analysis of Dyes and Inks;566
25.1;20.1 INTRODUCTION;566
25.2;20.2 DYES;568
25.3;20.3 INKS;570
25.4;20.4 CONCLUSION;594
25.5;References;594
26;Chapter 21 - Analysis of Dietary Supplements;600
26.1;21.1 INTRODUCTION;600
26.2;21.2 ANALYTICAL CHALLENGES;603
26.3;21.3 INSTRUMENTAL TLC;607
26.4;21.4 ANALYSIS OF BIOACTIVE INGREDIENTS;617
26.5;21.5 FINGERPRINTING AND DETECTION OF ADULTERANTS;638
26.6;21.6 CONCLUSIONS;641
26.7;References;642
27;Index;648

Leseproben

Chapter 1

Milestones, Core Concepts, and Contrasts

Colin F. Poole     Department of Chemistry, Wayne State University, Detroit, MI, USA

Abstract

Thin-layer chromatography and column chromatography are complementary separation techniques based on liquid chromatography. Although their application domains overlap, there is generally good reason to select one method over the other for particular applications. Capillary-controlled flow and the development mode are widely used in thin-layer chromatography. This restricts its kinetic performance compared to forced-flow techniques. Multiple development and multidimensional strategies increase the separation potential when using capillary-controlled flow. Incremental multiple development facilitates the use of solvent-strength gradients for the separation of mixtures with a wide range of retention properties. In this chapter, the general parameters used to describe separations in thin-layer chromatography are described and compared with the equivalent terms employed in column chromatography with a view to establishing the similarities and differences for the two techniques. This approach also affords general framework for method selection and to establish expectations.

Keywords

Capillary-controlled flow; Column chromatography; Forced flow; High-pressure liquid chromatography; History; Multiple development; Plate height; Plate number; Resolution; Retardation factor; Solvent-strength gradients; Spot capacity; Thin-layer chromatography; Two-dimensional chromatography

1.1. Introduction

Column chromatography and thin-layer chromatography are alternative formats for liquid chromatography [1]. Both formats exist as simple laboratory tools requiring little instrumentation and also as fully instrumental techniques. In both the cases, the stationary phase consists of a sorbent bed containing homogeneously packed particles or as a porous monolith. When movement of the mobile phase through the sorbent bed is controlled by capillary forces, the separation performance is suboptimal but requires little instrumentation affording a convenient and flexible arrangement for simple separations at the analytical or preparative scale. For faster separations, or separations with a higher peak capacity, a mechanism is required to enhance the mobile phase velocity. This requires instrumentation to pressurize the mobile phase and is the basis of high-pressure (or high-performance) liquid chromatography (HPLC) for columns and forced flow (or overpressured layer chromatography) for layers [2–4]. Although forced-flow instrumentation for thin-layer chromatography is commercially available, it is not in common use. Thus, whereas the practice of HPLC is a forced-flow technique, the practice of thin-layer chromatography is predominantly a capillary-controlled flow technique. In the latter case, instrumentation is required to optimize the various steps in the separation process and is referred to as high-performance thin-layer chromatography (HPTLC), or instrumental thin-layer chromatography, to distinguish the technique from conventional thin-layer chromatography (TLC) performed with much simpler equipment [5,6]. The general advantages of utilizing HPLC conditions versus conventional column chromatography are well known. The same argument cannot be made for conventional TLC versus HPTLC, and the general migration of separations from conventional TLC practices to HPTLC has not been universal. In fact, one might say that conventional TLC thrives in the laboratory environment as a quick, inexpensive, flexible, and portable method for surveying the composition of simple mixtures while only a few laboratories are equipped to perform more complex and quantitative analyses by HPTLC.

1.2. Milestones

The origins of thin-layer chromatography can be traced to the experiments on drop chromatography performed by Izmailov and Shraiber in the late 1930s [7]. From this beginning, thin-layer chromatography evolved into a fast and more powerful tool than gravity flow column chromatography for analytical separations. Thin-layer chromatography, as we know it today, was established in the 1950s due in large part to the efforts of Stahl and Kirchner on different continents. Their main contribution was the development of standardized materials and procedures that led to improved performance and reproducibility, as well as popularizing the technique by contributing many new applications [8]. At about the same time, commercialization of materials and devices commenced making the technique accessible to all laboratories. This ushered in the golden era of thin-layer chromatography where it quickly displaced paper chromatography as the main analytical liquid chromatographic method. By the 1970s, high-pressure liquid chromatography was becoming firmly established as an alternative approach for liquid chromatography and eventually grew to eclipse thin-layer chromatography for analytical applications. Thin-layer chromatography did not disappear in subsequent years but became less well known to those working in analytical laboratories where its strengths were often under appreciated. Developments continued in thin-layer chromatography as indicated by the time line Figure 1.1 [6,9].

First the development of high-performance thin-layer chromatography in the late 1970s is discussed. Layers coated with smaller particles of a narrow size distribution required the development of instruments for their convenient use. This was achieved by the early 1980s and so began the second era of thin-layer chromatography, known as modern or instrumental thin-layer chromatography. The evolutionary changes during this second era are captured in a series of books, which if ordered chronologically, represent the state-of-the-art at different times during this period to the present [5,10–15]. The main characteristic features of modern thin-layer chromatography are the use of fine particle layers for fast and efficient separations; sorbents with a wide range of sorption properties to optimize selectivity; the use of instrumentation for convenient and usually automated sample application, development and detection; and the accurate and precise in situ recording and quantification of chromatograms. Improvements in virtually all aspects of thin-layer chromatography continued over the next quarter century as indicated in Figure 1.1 and form the basis of subsequent chapters in this book. This period also marks the beginning of the philosophical division between conventional and high-performance thin-layer chromatography that has not been crossed by all those who use thin-layer chromatography. Expectations in terms of performance, ease of use, and quantitative information from the two approaches to thin-layer chromatography are truly opposite (see Section 1.1). As an example of expectations for a separation by modern thin-layer chromatography, see the chromatogram in Figure 1.2 for structurally similar ethyl estrogens (steroids used for birth control) [2]. Because of the small structural differences for these compounds, a high selectivity is required for their separation. Baseline separation is obtained with a short migration distance typical of fine particle layers and scanning densitometry provides a conventional record of the separation in the form of a chromatogram, as well as quantification of individual steroids after calibration. The quantitative results for tablet analysis are as accurate and precise as other chromatographic methods and the method is suitable for high-throughput routine tablet conformity analysis in which sample preparation requires no more than dissolution and filtration. Some specific reasons for choosing thin-layer chromatography for quantitative analysis are outlined in the next section.

FIGURE 1.1 Time line depicting important developments in the evolution of modern thin-layer chromatography. FFD=forced-flow development in an overpressured development chamber; AMD=automated multiple development chamber; AMC=automatic development chamber; and PPEC=pressurized planar electrochromatography.

FIGURE 1.2 Separation of ethynyl steroids by modern thin-layer chromatography. Two 15min developments with the mobile phase hexane–chloroform–carbon tetrachloride–ethanol (7:18:22:1) on a silica gel HPTLC plate. The chromatogram was recorded by scanning densitometry at 220nm. Reproduced with permission from Ref. [2].

1.3. Attributes of a Planar Format

Columns afford a better arrangement for operation at high pressures and for variation of the separation conditions by the control of external parameters. The thin-layer format provides a better arrangement for high sample throughput, flexible detection strategies, and a greater tolerance of samples with a high-matrix burden [2,16]. The throughput advantage is a consequence of the possibility of separating multiple samples in parallel with each sample occupying a single lane (or track) on the layer and several samples assigned to different lanes for simultaneous separation. Column chromatography is inherently a sequential separation process in which the separation time for a group of samples is the product of the...