Course Contents

CHROMATOGRAPHY Introduction to chromatography, Definition of chromatography, Types of chromatography, Theoretical principles underlying chromatographic techniques, Theories of chromatography; Plate theory, Rate theory, Zone shape; Standard deviation as a measure of zone breath, Calculation of H and N from zone breath, Experimental evolution of N and H, Sources of zone broadening; Eddy diffusion, Longitudinal diffusion, Non-equilibrium mass transfer, Net effect of the zone broadening processes, Separation on columns; Rate of solute migration, Relationship between retention time and partition coefficient, The Capacity factor, column resolution, Optimization of column performance, Column efficiency, Variation in the capacity factor, variation in the Selectivity factor, Development of the chromatography; Frontal analysis, Elution analysis, Displacement analysis, Selection of a chromatography system, Qualitative and quantitative analysis by chromatography; Qualitative analysis, Quantitative analysis; Analysis band on peak areas, Calibration with standards, The internal standard method, The area normalization method. THIN- LAYER CHROMATOGRAPHY Introduction, Superiority of TLC over other chromatographic techniques, Experimental techniques; Coating material, Preparation of thin layer plates, Activation of adsorbent, Purification of silica gel, Sample application, development tank, Solvent system, Detection of components, Evolution of the chromatogram, Direct method, Indirect method, Applications of TLC, Applications of TLC for separation of Inorganic Ions, Separation of amino acids, Separation of vitamins by TLC, Applications of TLC in quantitative analysis, Applications of some other forms of TLC; Adsorption TLC, Ion exchange TLC, Parathion TLCL, Reversed phased parathion TLC, In reversed phase parathion TLC, Limitation, Scope, High performance Thin layer chromatography. GEL PERMEATION CHROMATOGRAPHY Introduction, Principle, Materials, Gel preparation, Column packing and detectors, Applications, Advantage of Gel chromatography. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY Introduction, Principle, Instrumentation, Apparatus and materials; The column, Column packing, Column packing procedure, Chromatographic solvent, Pumping system, Detector system, Practical procedure, Column efficiency and selectivity, Comparison of High performance liquid chromatography and Gas chromatography, Applications, HPLC adsorption chromatography, HPLC partition chromatography. FAST PROTEIN LIQUID CHROMATOGRAPHY Introduction, Theory of FPLC, instrumentation, working techniques, applications

Course Synopsis

This module will explore the tenets of modern chromatography and explain the underlying theories and applications.

Course Learning Outcomes

Having successfully completed this module students will be able to: • Have raised awareness of chromatographic applications areas • Explain the principles of the most important TLC, HPLC, Gel Permetion Chromatography • Determine appropriate chromatographic technique and approach for analysis • Comprehend the optimization of chromatographic methods • Have raised awareness of chromatographic applications areas • Critically evaluate the quality of acquired data • Evaluate strengths and limitations of the most important chromatographic separation and detection methods in relation to the properties of the sample and of the analysis task