Transcript Diapositive 1 - Jagiellonian University

An introduction to chromatography
Objective of chromatographic methods
To identify the compounds of a mixture = qualitative analysis
To quantify these compounds
To retrieve the separated solutes (preparative techniques)
Spectroscopic methods (UV-vis, IR, RMN, MS) can also be used directly for
mixtures of compounds but are limited to 2 or 3 analytes at a time.
The objective of chromatographic methods is to separate the
compounds present in a mixture.
It is then possible to identify and quantify them.
The three steps of chromatography
Introduction of the sample in the separative device
(the sample must have been prepared for this purpose)
Separation with stationary and mobile phases
Detection of the separated species getting out of the
chromatographic system
The three steps of chromatography
A chromatogram is produced:
Solute 1
Solute 2
Analysis time
The three steps of chromatography
Basic principle of chromatographic techniques:
Different affinity of the solutes for two non-miscible phases
Stationary phase (SP): Solid or Liquid
Mobile phase (MP): Gaseous, Liquid or Supercritical
The three steps of chromatography
Pressure (MPa)
Supercritical fluid
9
8
31°C
* 7.3 MPa
7
Liquid
6
Gas
10
20
30
40
Temperature °C
State diagram for carbon dioxide
Homogeneous phase separation methods
NON CHROMATOGRAPHY
Driving force
Method
Centrifugal force
Centrifugation
Electrical
Electrophoresis
Electrodialysis
Concentration gradient
Dialysis, Osmosis
Pressure
Microfiltration, Ultrafiltration
Reversed osmosis
Field Flow Fractionation
Principles of separation used in chromatography
Property
Technique
Example Application
Molecular size
Size Exclusion Chromatography
Polymers
Volatility
Gas Phase Chromatography
Perfumes
Charge
Ion Exchange Chromatography
Amino acids
Solubility
Partition Liquid Chromatography
Any solute
Polarity
Adsorption Liquid Chromatography Any solute
Thin Layer Chromatography
Plant extracts
Specific affinity
Affinity Chromatography
Proteins
Chirality
Chiral chromatography
Drugs, perfumes