Chromatographic Techniques


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.


Bartle KD and Myers P (2001) Capillary Electrochromatography. Cambridge, UK: The Royal Society of Chemistry. ISBN: 0‐85404‐530‐9.

Bhattacharyya L and Rohrer JS (2012) Applications of Ion Chromatography for Pharmaceutical and Biological Products. John Wiley & Sons. ISBN: 9780470467091.

Irvine GB (1998) Size‐exclusion high‐performance liquid chromatography of peptides: a review. Analytica Chimica Acta 352 (1‐3): 387–397.

Jennings W, Mittlefehldt E and Stremple P (1997) Analytical Gas Chromatography, 2nd edn. Academic Press. ISBN: 978‐0‐12‐384357‐9.

Lewis AC, Carslaw N, Marriott PJ, et al. (2000) A larger pool of ozone‐forming carbon compounds in urban atmospheres. Nature 405: 778–781.

Martin AJ and Synge RL (1941) A new form of chromatography employing two liquid phases. Biochemical Journal 35: 1358–1368.

Meyer V (1999) Practical High‐Performance Liquid Chromatography. Chichester, UK: Wiley.

Mohammad A, Fatima N, Ahmad J and Khan MAM (1993) Planar layer chromatography in the analysis of inorganic pollutants. Journal of Chromatography 642: 445–453.

Poole CF (2003) Thin‐layer chromatography: challenges and opportunities. Journal of Chromatography, A 1000: 63–984.

Poole CF (2012) Stationary phases for packed‐column supercritical fluid chromatography. Journal of Chromatography, A 1250: 157–171.

Skoog DA, Holler FJ and Nieman TA (1998) Principles of Instrumental Analysis. Orlando, FL: Harcourt Brace.

Tswett M (1906) Physikalisch‐Chemische Studien über das Chlorophyll. Die Adsorption. (Physical‐chemical studies of chlorophyll adsorption. Berichte der Deutschen botanischen Gesellschaft 24: 316–326.

Van der Horst A and Schoenmakers PJ (2003) Comprehensive two‐dimensional liquid chromatography of polymers. Journal of Chromatography, A 1000: 693–709.

Wu CS (ed.) (1995) Handbook of Size Exclusion Chromatography. New York: Marcel Dekker.

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.

Poole CF (1999) Planar chromatography at the turn of the century. Journal of Chromatography A 856: 399–427.

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.

Reid E (2013) Bioanalysis of Drugs and Metabolites, Especially Anti‐Inflammatory and Cardiovascular. New York: Springer‐Verlag.

Rudd PM, Guile GR, Kuster B, et al. (1997) Oligosaccharide sequencing technology. Nature 388: 205–207.

Touchstone JC and Levin S (eds) (1990) Planar Chromatography in the Life Sciences. Chichester, UK: Wiley.

Zhou ZL, Licklider LJ, Gygi SP and Reed R (2002) Comprehensive proteomic analysis of the human spliceosome. Nature 419: 182–185.

Contact Editor close
Submit a note to the editor about this article by filling in the form below.

* Required Field

How to Cite close
Lewis, Alastair C(Feb 2015) Chromatographic Techniques. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0002705.pub2]