Chromatography, Principle and its Applications

Chromatography is an important biophysical technique that enables the separation, identification, and purification of the components of a mixture for qualitative and quantitative analysis. The Russian botanist Mikhail Tswett coined the term chromatography in 1906. The first analytical use of chromatography was described by James and Martin in 1952, for the use of gas chromatography for the analysis of fatty acid mixtures. A wide range of chromatographic procedures makes use of differences in size, binding affinities, charge, and other properties to separate materials. It is a powerful separation tool that is used in all branches of science and is often the only means of separating components from complex mixtures.

Principle of Chromatography

Chromatography is based on the principle where molecules in mixture applied onto the surface or into the solid, and fluid stationary phase (stable phase) is separating from each other while moving with the aid of a mobile phase.

• The factors effective on this separation process include molecular characteristics related to adsorption (liquid-solid), partition (liquid-solid), and affinity or differences among their molecular weights.
• Because of these differences, some components of the mixture stay longer in the stationary phase, and they move slowly in the chromatography system, while others pass rapidly into the mobile phase, and leave the system faster.

Applications of Chromatography

Pharmaceutical sector

• To identify and analyse samples for the presence of trace elements or chemicals.
• Separation of compounds based on their molecular weight and element composition.
• Detects the unknown compounds and purity of mixture.
• In drug development.

Chemical industry

• In testing water samples and also checks air quality.
• HPLC and GC are very much used for detecting various contaminants such as polychlorinated biphenyl (PCBs) in pesticides and oils.
• In various life sciences applications

Food Industry

• In food spoilage and additive detection
• Determining the nutritional quality of food

Forensic Science

• In forensic pathology and crime scene testing like analysing blood and hair samples of crime place.

Molecular Biology Studies

• Various hyphenated techniques in chromatography such as EC-LC-MS are applied in the study of metabolomics and proteomics along with nucleic acid research.
• HPLC is used in Protein Separation like Insulin Purification, Plasma Fractionation, and Enzyme Purification and also in various departments like Fuel Industry, biotechnology, and biochemical processes.