2. Presentation on :
Partition Column Chromatography
Course code : ACCE3105
Course Title : Instrumental Method of analysis- I
Submitted to:
Rajib Chandra Das
Lecturer
Dept. of ACCE,NSTU
NOAKHALI SCIENCE & TECHNOLOGY UNIVERSITY
Department of Applied Chemistry & Chemical Engineering
Date of Submission: 11/07/18
2
Presented by
ASIF PAPPU
3. Partition Column
Chromatography
• Chromatography
• Partition chromatography
• Types of partition chromatography
• Principle of partition column chromatography
• Application of partition column chromatography
3
4. Partition Chromatography
• Method of separation in which the components present in the
mixture get distributed more likely into two liquid phases because of
differences in partition coefficients during the flow of mobile phase in
the chromatography column.
4
5. Partition Chromatography
• The most widely used type of HPLC is partition chromatography .
• The stationary phase is a second liquid that is immiscible with the
liquid mobile phase.
• Earlyr in PC used liquid liquid column but in modern LC systems
liquid bonded phase column is used.
• In liquid liquid chromatography the liquid was held in place by
physical adsorption but in liquid bonded phase column system,
attached by chemical bonding resulting in highly stable packing
insoluble in the mobile phase. Bonded - phase columns are also
compatible with gradient elution technique
5
6. Column for Bonded-Phase Chromatography
6
In maximum bonded phase packing
for partition chromatography are
prepared from rigid silica or silica
based compositions .This is uniform
porous and mechanically study.
Gradient elution – Mobile phase content changes
during elution
Isocratic elution – Mobile phase is all the time the same
during elution
7. 7
Difference Between Adsorption and Partition Chromatography
• Partition chromatography - separation on the stationary phase
occurs by partition due to differences in partition coefficients.
• Used for liquid-liquid or liquid-gas chromatography
• Adsorption Chromatography -relative differences in adsorption
of constituents of given sample. Because of differences in their
affinity towards stationary phase, the components of the mixture
adsorb with different rates.
• used only for solid-liquid or solid-gas chromatography.
8. Partition Chromatography Principle
• Separation of components of given sample occurs due to partition of
components between two liquid phases.
• Stationary phase is coated with a liquid which is immiscible in mobile
phase.
• Stationary phase immobilizes the liquid surface and makes it
stationary phase.
• The mobile phase passes over the stationary phase and separate out.
• The separation depends on the relative solubility in the stationary
liquid layer because of different partition coefficient, different
component of sample are separated.
8
11. Liquid - Liquid Chromatography
Partition or liquid-liquid chromatography (LLC): A powerful separation
technique which is used for the separation and analysis of acids and
proteins.
The basis of LLC is the distribution of sample molecules between two immiscible
liquid phases, a stationary phase and a mobile phase (Figure 1).
In conventional LLC, the stationary phase is mechanically held to a support by
adsorption.
Employs liquid mobile and stationary phases.
Uses small particles with molecules bonded to their surface to give a thin film that
has liquid like properties.
11
13. Application of LLC
• Separation of compounds containing metals.
• Determination of compounds in biological samples.
• Analysis of drugs in food products.
• Determination of compounds in environmental samples.
• Determination of compounds in biological samples.
• Separation of important organic chemical.
13
14. Partition Chromatography Applications
• used for final purification natural extracts, synthetic mixtures and
biological matrices.
• It is also used for fractionization of complex crude extracts eg.
Petroleum fractions
• Determination of water quality.
• Separation of aroma molecules of wine.
• Determination of pesticide residue.
14
