Liquid Chromatography


Liquid chromatography, with various modes, is a technique, which deals with the separation of molecules in samples by using specific solvents or buffers. Liquid chromatography or high‐pressure liquid (HPLC) is a popular quantitative analytical technique applied in many areas of chemical, biomedical and pharmaceutical sciences. HPLC is used for separation, detection and measurement of amino acids, proteins, lipids, carbohydrates, vitamins, trace elements and ions in various types of samples. It is also used for the analysis of polymorphisms in genes coding for specific proteins or enzymes. HPLC is widely used in diagnostic and research purposes in industry, hospitals, educational and research institutions. Liquid chromatography currently accounts for about 60% of separation technology applied in life sciences worldwide. HPLC analytical methods have high assay precision, specificity and accuracy.

Key Concepts

  • HPLC is an important analytical technology used worldwide.
  • HPLC is used for the analysis of micro‐ and macromolecules in human body fluids, water and food items for research and diagnostic purposes.
  • HPLC is also used for the analysis of polymorphisms in genes coding for the enzymes.

Keywords: HPLC; columns; detectors; analysis; solutes; solvents; buffers; adsorbents; counter‐ions

Figure 1. The column and the tubings in an HPLC system.
Figure 2. A typical HPLC system consisting of a degasser, a pump, mobile‐phase delivery system (isocratic or gradient), a detector (UV, fluorescence or electrochemical), auto‐sampler and integrator (computer‐aided). The column may be placed in an oven for constant temperature.
Figure 3. A typical chromatogram of a monobromobimane (MBrB)‐derivatised plasma sample of (a) a healthy control subject, (b) an epilepsy patient on anti epileptic drug and (c) a patient with kidney disease. Samples were analysed according to the method of Mansoor et al. (). Cys, cysteine; CG, cysteinylglycine; HCY, homocysteine. Column, 150 mm × 4.6 mm i.d.; stationary phase, particles ODS 3; mobile phase contains ion‐pairing agent tetrabutylammonium phosphate; column temperature 25°C.
Figure 4. A standard curve for cysteine.


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Further reading

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How to Cite close
Mansoor, Mohammad Azam(Jun 2015) Liquid Chromatography. In: eLS. John Wiley & Sons Ltd, Chichester. [doi: 10.1002/9780470015902.a0002679.pub3]