Chromatography
Applications
What is Chromatography?
Different types
Allows the separation, identification and determination of chemical components in complex mixtures.
No other separation method is so powerful and so generally applicable.
Chromatographies are processes that encompass several separation techniques, based on physical properties of certain materials
In interaction with substances or mixtures of substances, which are related to their chemical properties, allow the decomposition of a mixture and analyze its constituents.
With the development of technology, chromatographic techniques diversified and significantly improved their ability to resolve mixtures of different natures
Adsorption Chromatography
Thin Layer Chromatography
Column Chromatography
Ion-exchange chromatography
Affinity chromatography
Paper chromatography
Gas chromatography
Hydrophobic interaction chromatography
This technique is used for the purification of biomolecules. On a column (stationary phase) firstly the sample to be separated
Their flow through inside column material placed on a fiberglass support is ensured.
The samples are accumulated at the bottom of the device in a tme-, and volume-dependent manner
based on electrostatic interactions between charged protein groups, and solid support material (matrix).
Matrix has an ion load opposite to that of the protein to be separated, and the affinity of the protein to the column is achieved with ionic ties.
Proteins are separated from the column either by changing pH, concentration of ion salts or ionic strength of the buffer solution
positively charged ion- exchange matrices are called anion-exchange matrices, and adsorb negatively charged proteins.
While matrices bound with negatively charged groups are known as cation-exchange matrices, and adsorb positively charged proteins
this chromatography technique is used for the purification of enzymes, hormones, antibodies, nucleic acids, and specific proteins
A ligand which can make a complex with specific protein (dextran, polyacrylamide, cellulose etc) binds the filling material of the column.
The specific protein which makes a complex with the ligand is attached to the solid support (matrix), and retained in the column, while free proteins leave the column.
Then the bound protein leaves the column by means of changing its ionic strength through alteration of pH or addition of a salt solution
consists of a layer of cellulose highly saturated with water.
Mobile phase consists of an appropriate fluid placed in a developing tank
In this method a thick filter paper comprised the support, and water drops settled in its pores made up the stationary “liquid phase.”
Paper chromatography is a “liquid-liquid” chromatography
“solid-liquid adsorption” chromatography. In this method stationary phase is a solid adsorbent substance coated on glass plates.
As adsorbent material all solid substances used. in column chromatography (alumina, silica gel, cellulose) can be utilized
the mobile phase travels upward through the stationary phase The solvent travels up the thin plate soaked with the solvent by means of capillary action.
it also drives the mixture priorly dropped on the lower parts of the plate with a pipette upwards with different flow rates. Thus the separation of analytes is achieved.
stationary phase is a column which is placed in the device, and contains a liquid stationary phase which is adsorbed onto the surface of an inert solid. Gas chromatography is a “gas-liquid” chromatography.
ts carrier phase consists of gases as He or N2. Mobile phase which is an inert gas is passed through a column under high pressure.
The sample to be analyzed is vaporized, and enters into a gaseous mobile phase phase. The components contained in the sample are dispersed between mobile phase, and stationary phase on the solid support.
Gas chromatography is a simple, multifaceted, highly sensitive, and rapidly applied technique for the extremely excellent separation of very minute molecules.
In this method the adsorbents prepared as column material for the ligand binding in affinity chromatography are used.
HIC technique is based on hydrophobic interactions between side chains bound to chromatography matrix
different compounds are adsorbed on the adsorbent to different degrees based on the absorptivity of the component.
a mobile phase is made to move over a stationary phase, thus carrying the components with higher absorptivity to a lower distance than that with lower absorptivity
The main types of chromatographic techniques that are used in industries are given as under.
Examining reaction mixtures
Quality control
Identifying drug residues in food
Food industry
Forensic science
Food analysis
application of science principles and methods to support legal decision-making in matters of criminal and civil law
During criminal investigation in particular, it is governed by the legal standards of admissible evidence and criminal procedure.
Paper chromatography has traditionally been used to analyse food colours in ice creams, sweets, drinks and beverages, jams and jellies.
It is very helpful in separating compound mixtures.
TLC can be used to study if a reaction is complete. At the beginning of a reaction
as a reaction monitoring option, the rapid methods using spectroscopic techniques are limiting the paper chromatography application
uality control. Manufacturing relies on gas chromatography for quality control, companies that produce cars, chemicals, and pharmaceuticals, in particular
Quality control is a crucial application of gas chromatography within the food industry.
Spoilage detection is one of the most notable uses of chromatography in the food industry
Food and Beverage. Quality control within the food and beverage industry can be enacted through chromatography
Food analysis and quality control are one of the most common applications for gas chromatography due to their component quantification, accuracy, and speedy
Gas chromatography is vital to the food industry for ensuring the safety of food products, preventing contaminated products