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Chromatography

Chromatography is used to separate and analyze complex mixtures by passing a sample through a stationary and mobile phase. There are four principles of chromatography including molecular exclusion, charge properties, partitioning, and adsorption. Ion-exchange chromatography separates ions and polar molecules based on their charge, and is often used in protein purification and water analysis.

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28 views

Chromatography

Chromatography is used to separate and analyze complex mixtures by passing a sample through a stationary and mobile phase. There are four principles of chromatography including molecular exclusion, charge properties, partitioning, and adsorption. Ion-exchange chromatography separates ions and polar molecules based on their charge, and is often used in protein purification and water analysis.

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ulfh08152
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© Attribution Non-Commercial (BY-NC)
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Download as PPT, PDF, TXT or read online on Scribd
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CHROMATOGRAPHY -used to separate and or to analyze complex mixtures -It involves passing a sample (the analyte) in the "mobile

phase", often in a stream of solvent, through the "stationary phase

Stationary phase
- Part of the chromatographic system through which the

mobile phase flows where distribution of the solutes between the phases occurs. - May consists of particles (porous or solid), the walls of a tube (capillary) or a fibrous material (paper).
Mobile phase
- Part of the chromatographic system which carries the

solutes through the stationary phase.

4 principles of chromatography:
1) Molecular exclusion (Gel permeation)
- based on molecular size of the solute molecule - the solute with small size can penetrate the capillaries of the stationary phase and will be retained - the molecule with larger size will be excluded and pass through the flow

2) Charge properties -based on charge of the molecule -the positive charge will go to negative charge -get two layer -Ex: Ion exchanger chromatography.

3) Partitioning - Separation based on the relative solubility of each solute in the stationary phase

4) Adsorption - Separation based on the relative attraction of the solute molecules to the active surface groups resulted from nonuniform distribution of charges - dipole

Ion-exchanger chromatography -is a process that allows the separation of ions and polar molecules based on the charge properties of the molecules. -It can be used for almost any kind of charged molecule including large proteins, small nucleotides and amino acids. -The solution to be injected is usually called a sample, and the individually separated components are called analytes. -It is often used in protein purification, water analysis, and quality control.

Ion exchanger chromatography


-Cation exchange chromatography -Anion exchange chromatography

CATION EXCHANGE CHROMATOGRAPHY - Retains positively charged cations because the stationary phase displays a negatively charged functional group such as phosphoric acid

ANION EXCHANGE CHROMATOGRAPHY -Retains negatively charged anions using positively charged functional group.

Ion exchanger chromatography

-THE END-

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