Advances in Various Chromatographic Techniques

Chromatography is the method of separation of compounds from a mixture. The technique is both analytical and preparative and is employed widely in laboratory as well as industrial standards. More than 60% of chemical analysis all over the world is done with chromatography or a variation thereon. Chromatography is a physical technique applied from basic organic chemistry to Forensic sciences. Some common applications include detection of cocaine in urine, PCB’s in fish, alcohol in blood, and lead in water. Chromatography is of various types- Paper chromatography, displacement chromatography, thin layer chromatography, supercritical chromatography, Column fluid chromatography, gas chromatography, expanded bed adsorption chromatography, liquid chromatography in various combinations.

The increasing demand for monoclonal antibodies is driving the chromatography resin market globally. Chromatographic resins are used in separation and purification of proteins and other biomolecules in pharmaceuticals, food production, biotechnology, water and environmental analysis. Pharma and biotech industries are the largest consumer of chromatography resins followed by food production. North America has the highest market share of chromatography resins in terms of both consumption and revenue followed by Europe. The income from sales of resins in food production industry in North America is expected to record highest CAGR in the next five years. However, in Asian countries the chromatography resin consumption shows huge potential to grow in the pharmaceutical sector. The raising biosimilars market in emerging economies such as India, South Korea and China is expected to drive the chromatography resins market in the coming decade.

Column chromatography in chemistry is a method used to purify individual chemical compounds from mixtures of compounds. It is commonly used for preparative applications on scales ranging from micrograms to kilograms. The advantage of column chromatography is the relatively low cost and disposability of the stationary phase used in the process. Paper chromatography is a technique that involves placing a small dot or line of sample solution onto a strip of polar cellulose chromatography paper. The paper is placed in a glass chamber with a shallow layer of solvent and sealed. As the solvent moves through the paper, it contacts the sample mixture, which starts to rise up the paper with the solvent. Thin layer chromatography (TLC) is a widely employed technique which involves a stationary phase of a thin layer of adsorbent like alumina, silica gel, or cellulose on a flat, inert layer of substrate. TLC has the advantage of better separations, faster runs, and the choice of different adsorbents. Better resolution and quantification can be achieved with high-performance TLC.

Displacement chromatography is a chromatography technique in which a sample is placed onto the head of the column and is then displaced by a solute that is more strongly adsorbed than the components of the original mixture. As a result is that the components are resolved into consecutive rectangular zones of highly concentrated pure substances rather than solvent-separated peaks. It is primarily a preparative technique. Higher product concentration, higher purity, and increased throughput are obtained compared to other modes of chromatography.

Gas chromatography (GC) is a common type of chromatography used in analytical chemistry for separating and analysing compounds that can be vaporized without decomposition. In GC, the mobile phase (or "moving phase") is a carrier gas, commonly an inert gas such as helium or an unreactive gas such as nitrogen. Stationary phase is a microscopic layer of liquid or polymer on an inert solid support, within a glass or metal tubing called a column and the instrument used to perform gas chromatography is called a gas chromatograph (or "aerograph", "gas separator"). The gaseous compounds being analysed interact with the walls of the column coated with a stationary phase.

Supercritical fluid chromatography (SFC) is a separation technique in which the mobile phase is a fluid above and relatively close to its critical temperature and pressure. SFC typically utilizes carbon dioxide as the mobile phase; therefore the entire chromatographic flow path must be pressurized. Supercritical phase represents a state in which liquid and gas properties combine, Supercritical fluid chromatography is sometimes called convergence chromatography. Expanded Bed Adsorption (EBA) Chromatographic Separation captures a target protein from a crude feed stream when it passes through a chromatography column system containing adsorbent beads. Using this technique the unprocessed crude compound can be treated directly in the chromatographic column, avoiding clarification and pre-treatment steps. Supercritical fluid chromatography is a separation technique in which the mobile phase is a fluid above and relatively close to its critical temperature and pressure.

The basic chromatography technique includes a stationary phase and a mobile phase. The mobile phase carries the components to be separated through the stationary phase. The components are thus separated based on their relative affinity with the stationary phase. Additionally, preconditioning helps in better elution.

  • Column chromatography
  • Paper chromatography
  • Thin layer chromatography
  • Displacement chromatography
  • Gas chromatography
  • Supercritical fluid chromatography
  • Capillary electrophoresis
  • Expanded bed adsorption (EBA) chromatography

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