THEORY

HPLC (HIGH PERFORMANCE LIQUID CHROMATOGRAPHY)

In biochemistry, some of the main aims are to understand and describe the structural basis of biological molecules as they relate to function. The diversity of life is reflected by the enormous number of molecules found in living organisms. If these molecules are to be studied they must be purified. One of the biggest breakthroughs in biochemistry was the introduction of chromatography which made separation of molecules possible in an fast and efficient manner. Chromatography is the science of separation which uses a diverse group of methods to separate closely related components of complex mixtures.It ıs a physical separation of two or more compounds based on their differential distribution between two phases.

There are millions of proteins found in living organisms which makes the isolation of a single protein molecule a tedious and time consuming task. Protein molecules are separated according to their physical properties such as their size, shape, charge, hydrophobicity, and affinity for other molecules. The term High Performance Liquid Chromatography (HPLC) was coined to describe the separation of molecules under high pressure in a stainless steel column filled with a matrix. In HPLC, both pump control and data acquisition can be under the control of the same computer.

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