Different Types of chromatography techniques for Various Industries.
In Industries pertaining to the synthesis of bio-molecules require a special technique which does not denature the characteristics of the bio-molecules during Separation and Purification. Column Chromatography is one such technique having different modes which are used on a large scale in Industrial level as it is bio-compatible and requires relatively short time for separation and purification. The Column Chromatography Principle is mainly based on size, Charge, and Hydrophobicity of the sample which in turn lead to various Chromatographic techniques such as-Size exclusion chromatography or Gel permeation chromatography, Adsorption chromatography, Ion exchange chromatography and Affinity Chromatography. Each technique is based on the biological interaction between the sample bio-molecules and the packing material such as Alumina or Silica gel.
Types of chromatography and their applications
Size exclusion chromatography:
is also known as molecular sieve chromatography separates analyses according to their size as they pass on through the stationary phase namely Silica which is porous in nature and acts as a hydrophilic surface.
the stationary phase is considered as the Adsorbent which is used in separating Non-volatile mixtures. In this method, the Adsorbent used can be Silica, Alumina or Cellulose.
Ion exchange chromatography
is used in the separation of polar molecules and ions such as proteins, amino acids containing both positive and negative charged chemical groups. In this method, the stationary phase used is slurry of either Silica or Alumina.
Affinity chromatography is used for the separation of recombinant proteins and enzymes and here the stationary phase act as a Support medium.
Chromatography techniques are widely used-
- In Biochemical research applications to separate and identify the compound of biological origin,
- In Petroleum industries to control the production of saturated carbons,
- In Chemical Industries to purify the chemicals,
- In Sugar factories to purify sugar from Molasses.
For any technique, if the packing material employed is Silica gel than the Silica gel column chromatography Procedure mainly depends on packing the column without cracks and air bubbles as it leads to poor separation and purification. Whereas if the packing material used in these techniques is Alumina Column Methanol, it contains Methanol, which is considered as a volatile eluting solvent and therefore used in Alumina Column Chromatography. Apart, from methanol to improve the effectiveness of purification and to adjust the alkalinity it is treated with either dilute acid or base considering the components nature of the Column. After adjusting the alkalinity of Alumina it is characterized as Neutral, Acidic and Basic Alumina. Among this Neutral Alumina is used in chromatographic cleanup procedures and also for the separation of Aliphatic, Aromatic and Polar fractions. Purification of free base Porphyrins and Metalloporphyrins is also performed using Neutral Alumina. When it is used as a stationary phase in the Column, for the preparative separation of the mixture having basic properties, it does not require a mobile phase modifier.
Uses of Chromatography in medicine
Lack of efficiency of a medicine results in lethal accidents and has become a major concern. Therefore Chromatography opts as a detection tool in identifying and separating the desired medicine from the contaminants. The following are some of the medicines that are developed using Chromatography techniques-
- Synthesis of Potential Anti Pertussis medicine involves Chromatography technique.
- Separation of enantiomers which are useful in drug discovery is progressed through different Chromatography techniques.
- Chromatography is also applied in the batch Fermentation process to determine the improvement of the process during antibiotic production.
- Separation often different H1-Antihistamines which were structurally related was achieved by Chromatography, impregnating Silica gel with transition metal ions.
- Purification of Proteins and Peptide hormones is attained either by Ion-exchange Chromatography or Gel Permeation Chromatography.
- By Size exclusion chromatography using Alumina nanoparticles size separation of DNA molecules was achieved.
Application of Chromatography in Chemistry
In chemistry one of the major aspects is to analyze the residue of all major classes which include Food, water, and air. It is mainly essential to emphasize the quality and safety of these classes or else lead to adverse effect on Human health. Therefore, this analysis of residues is accomplished by different Chromatography Techniques. The following are some of the techniques used in chemistry to separate and analyze the components which are essential in day to day life.
- Separation of Proteins and small Oligonucleotides made easy by using Ion-exchange chromatography.
- It is also used for the separation of Aldehydes, Ketones, Quinones, Esters, Lactones, and Glucosides.
- Environmental agency as made Chromatography a method to test the quality of the Drinking water.
- It is also used to monitor the air quality.
- It is employed in the separation of organic as well as Inorganic compounds.
- It is an indispensable method to study the heavy oils containing hydrocarbon group.
- Analysis of pesticide residues on fruits and vegetables is also carried by this analytical technique.