Etiket Arşivleri: High Performance Liquid Chromatography

High Performance Liquid Chromatography ( Dr.Hüseyin BOZKURT )

High Performance Liquid Chromatography (HPLC) is one mode of chromatography, the most widely used analytical technique. Chromatographic processes can be defined as separation techniques involving mass-transfer between stationary and mobile phases. Present day liquid chromatography that generally utilizes very small packing particles and a relatively high pressure is referred to as high performance liquid chromatography (HPLC).

HPLC utilizes a liquid mobile phase to separate the components of a mixture. These components are first dissolved in a solvent, and then forced to flow through a chromatographic column under high pressure. In the column, the mixture is resolved into its components. The amount of resolution is important, and is dependent upon the extent of interaction between the solute componentsand the stationary phase. The stationary phase is defined as the immobile packing material in the column. The interaction of the solute with mobile and stationary phases can be manipulated through different choices of both solvents and stationary phases. As a result, HPLC acquires a high degree of versatility not found in other chromatographic systems and has the ability to easily separate a wide variety of chemical mixtures.

HPLC ( High Performance Liquid Chromatography )

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.