Chromatography Flashcards
Most frequently used analytical technique in pharmaceutical analysis
Chromatography
Technique to separate plant pigments by passing solutions through glass colums packed with finely divided calcium carbonate
Chromatography
Chromatography is widely used for
Identification, separation, determination of the chemical components in complex mixtures
Separate & identify components of a mixture based on the differential affinities of the solutes between 2 phases.
Chromatography
Porous or finely divided solid, or a liquid that has been coated in a thin layer on an inert supporting material. A fixed bed core of large surface area.
Stationary Phase
A pure liquid or a mixture of solutions (eg, buffers) or it may be gas (pure or homogeneous mixture) or supercritical liquid
A fluid w/c moves through/ or over the surface/ of the stationary phase.
Mobile Phase
Addition of a flowing solvent on the stationary phase to wash out, purify, or separate material
Elution
Principal objectives obtainable through the use of Chromatography are:
• Resolution of mixtures into constituent parts.
• Determination of homogeneity.
• Comparison of substances suspected of being identical.
• Purification Concentration of substances from dilute solutions.
• Identification & control of technical products.
• Quantitative separation from complex mixtures.
• Indication of molecular structure.
Basic Principles of Separation/Equilibration Process
Partition
Adsorption
Affinity
Moelcular Exclusion/Pore penetration
Ion Exchange
The components distribute b/n two phases through a combination of sorption nd desorption process
Adsorption chromatography
Mixtures of solutes are spearated according to the relative tendencies of their components to partition between 2 immiscible gas or L-G
Partition Chromatography
Adsorption Chromatograpjy
Stationary Phase: Solid
Mobile Phase: Liquid/Gas
Examples: TLC, CC, HPC
Partition Chromatography
Stationary Phase: Liquid/Gas
Mobile Phase: Liquid
Examples: PC, TLC, HPLC
The mobile phase passes over the surface, the ionic solutes are retained by forming electrostatic chemical bonds with the functional group.
Ion Exchange Chromatography
Ion Exchange Chromatography
Stationary Phase: Polymeric matrix, Resin/Ionic functional groups
Mobile Phase: Liquid
Examples: IEC, HPLC
Molecular Exclusion Chromatography aka
Pore Penetration
Size Exclusion
Gel Filtration/ Gel permeation/ Gel chromatography
Used to separate groups of solutes based on effective size in solution/ based on MW.
Molecular/ size exclusion Chromatography
Use of Molecular Exclusion Chromatography
Preparative separations of macromolecules of biological origin
Purification of synthetic organic polymers
Molecular Chromatography
Stationary Phase: Polymeric subs containing pores of molecular dimensions
Mobile Phase: Liquid/Gas
Examples: MEC, HPLC
utilizes highly specific interactions between one kind of solute molecul and a second molecule covalently attached to stationary phase
Affinity Chromatography
Affinity Chromatography
Stationary Phase: Solid, affinity ligand/antibody enzyme
Mobile Phase: Liquid
Examples: DNA Affinity
A graphical representation of the detector response, concn of analyte in the effluent; A measure of effleunt concn vs volume/time
Chromatogram
Fresh portions of solvent added on the top of the comlumn to bring about separation of the componenets as each drug progresses down the column
Developer
Washing out, purify, or separate materials; or to remove one substance to another, usually an absorbed material from an adsorbent surface by washing it out by a solvent.
Elution
