Items related to Thin-Layer Chromatography: A Laboratory Handbook
Synopsis
General Section.- A. History of the Development of Thin-Layer Chromatography.- B. Instruments used in Thin-Layer Chromatography and their Operation.- I. The application of thin layers to carrier plates.- Thin-layer spreader.- a) Aligning tray.- b) Positioning of the spreader.- c) Adjustment of layer thickness.- d) Filling of spreader and application.- e) Cleaning and storage of spreader.- II. Drying, storage and handling of TLC-plates.- III. Preparation of TLC-plates for use in chromatography.- a) Checking dried thin layers.- b) Stripping of thin-layer edge.- c) Marking the layer.- d) Instruments for application of spots.- e) Streak application of larger quantities of mixture for micro-preparative thin-layer chromatography.- IV. Separation chambers and conditions of saturation.- 1. Saturation and edge effects.- 2. Assembly of chambers (temperature, light, protection against oxidation).- 3. Chambers for ascending development.- a) Rectangular trough chamber.- b) S-Chamber system.- 4. Equipment for descending development.- 5. Equipment for horizontal development.- a) Circular technique.- b) Horizontal method in closed tank.- c) Continuous flow technique (BN-Chamber).- 6. Apparatus for electrophoresis and ionophoresis in thin layers of adsorbent.- V. Spraying equipment and fume hoods.- VI. Standard conditions in thin-layer chromatography.- VII. Basic equipment for use in thin-layer chromatography.- C. Coating Materials for Thin-Layer Chromatography.- Further properties of adsorbents.- Additional adsorbents and adsorbent combinations.- Storage and treatment of adsorbents.- D. Special Techniques.- 1. Continuous flow, and multiple development techniques.- 2. Wedged-tip technique.- 3. Two-dimensional separation, SRS-technique.- 4. Variation in the separation characteristics of a layer.- Bibliography to Chapters A-D, General Section.- E. Documentation of Thin-Layer Chromatograms.- F. Quantitative Evaluation of Thin-Layer Chromatograms.- I. Determination without extraction of separated substances from the chromatogram (Method I).- 1. Method of visual comparison.- 2. Evaluation by photographic methods.- 3. Photo-densitometric determination after staining.- 4. Autoradiographic evaluation.- II. Determination of separated substances after extraction (Method II).- 1. Location by color or fluorescence.- 2. Use of fluorescent layers.- 3. Staining of separated substances before extraction.- 4. Other methods of locating separated substances.- Bibliography to Chapters E and F, General Section.- G. Isotope Techniques.- I. Layers, solvents, and chemical methods of detection.- II. Methods of detecting radiation.- 1. Autoradiography.- 2. Counting tubes and scintillation counters.- III. Preparation of radioactively labelled substances.- IV. Isolation of radioactive compounds by thin-layer chromatography.- V. Analysis by means of radioisotopes.- 1. Indicator analysis.- 2. Isotope dilution analysis.- 3. Activation analysis.- 4. Isotopic derivative method.- a) Fractionation before radioactive labelling.- b) Separation of radioactive derivatives.- c) Fractionation after adding a radioactive derivative to the mixture of non-labelled derivatives.- d) Separation after adding an inactive derivative to the mixture of radioactively labelled derivatives of the compound to be determined.- e) Application of two different radioactive isotopes.- VI. Procedures for radioactive labelling.- 1. Esterification of acids with diazomethane, C14H2N2.- 2. Acetylation of alcohols with acetic anhydride (C14H3CO)2O or (CH33CO)2O.- VII. Application of thin-layer chromatography in chemical and biochemical investigations with radioisotopes.- Bibliography to Chapter G, Isotope Techniques.- H. Theoretical Aspects of Thin-Layer Chromatography.- General Remarks.- I. General theory of chromatography.- 1. An introductory experiment.- 2. Another experiment.- 3. The model.- 4. The chromatographic column.- a) Comparison with the model.- b) Developing a chromatogram.- c) Elution.- d) Estimat
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