Chromatographic Techniques

Abstract

Chromatography achieves the physical separation of chemical components in a mixture for either analytical or preparative purposes, based on the unique degree of interaction between each component and the separating medium. Chromatography can be used to isolate a wide range of different chemicals, from permanent gases to macromolecules, metals, ions and organic and inorganic compounds. Chromatography uses gases, liquids or supercritical fluids as the mobile phase to carry analytes through the separating medium, commonly referred to as the column. In the case of liquid chromatography, the column is often a packed tube, and in gas chromatography a wall‐coated capillary. Chromatography is used in many different application areas and is one of the most commonly used laboratory techniques. Chromatography is often coupled with mass spectrometry as a detector.

Key Concepts

  • Materials found in nature and from man's activities are often highly complex mixtures of different chemicals; isolating and quantifying individual components in such mixtures is a very common measurement requirement.
  • Chromatography is an analytical method in which a physical separation of individual components in a mixture is achieved.
  • A sample mixture is solvated in a fluid, which is used to carry it through a separating medium often referred to as a column. The solvating fluid is referred to as the mobile phase and the separating medium as the stationary phase.
  • Each analyte may interact (e.g. by adsorption) to a unique degree with the stationary phase, resulting in the physical separation of the mixture into its component parts.
  • The separating medium is often found in the form of a packed column of stationary‐phase material, open tubular column with a wall coating or as a planar coated plate.
  • Chromatographic methods are very commonly coupled with detectors such as mass spectrometry, which allows for the quantification and identification of analytes as they elute from the column.

Keywords: liquid chromatography; gas chromatography; ion‐exchange chromatography; size‐exclusion chromatography; complex mixtures

Figure 1. Schematic representation of the chromatographic process as it occurs in packed‐column elution chromatography. Two components (marked as black and white spots) are introduced together as a mixture at the start of the column. As they pass through the packed bed of stationary phase particles, they are separated based on their interactions with the stationary phase. Above the column is the resulting chromatogram that would be obtained. The size/height of each peak is a function of the original analyte concentration and specific detector response to that compound.
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Further Reading

Chester TL and Parcher JF (2001) Blurring the boundaries. Science 291: 502–503.

McNair H and Miller J (eds) (1997) Basic Gas Chromatography. Chichester, UK: Wiley.

Mondello L (ed.) (2011) Comprehensive Chromatography in Combination with Mass Spectrometry. Chichester, UK: Wiley.

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Poole CF and Dias NC (2000) Practitioner's guide to method development in thin‐layer chromatography. Journal of Chromatography 892: 123–142.

Poole SK, Kollie TO and Poole CF (1994) Influence of temperature on the mechanism by which compounds are retained in gas–liquid chromatography. Journal of Chromatography A 664: 229–251.

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Lewis, Alastair C(Feb 2015) Chromatographic Techniques. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0002705.pub2]