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High performance liquid chromatography (HPLC):
Introduction:
All chromatographic systems consist of the stationary phase, which may be a solid, gel, liquid or a solid/liquid mixture that is immobilised, and the mobile phase, which may be liquid or gaseous and which flows over or through the stationary phase. The choice of stationary phase is made so that the compounds to be separated have different distribution coefficients, which describes the way in which a compound distributes itself between 2 immiscible phases.
High performance liquid chromatography (HPLC) is a form of column chromatography used frequently in biochemistry and analytical chemistry. HPLC is used to separate components of a mixture based on a variety of chemical interactions between the substance being analyzed (analyte) and the chromatography column.
In isocratic HPLC the analyte is forced through a column of the stationary phase by a liquid at high pressure. Use of pressure gives the components less time to diffuse within the column, leading to improved resolution in the resulting chromatogram. Solvents used include any miscible combination of water or various organic liquids (the most common are methanol and acetonitrile). Water may contain buffers or salts to assist in the separation of the analyte components.
The internal standard is a compound that has physical properties as similar as possible to the test analytes and which chromatographs near to, but distinct from, them.
High performance liquid chromatography (HPLC):
Introduction:
All chromatographic systems consist of the stationary phase, which may be a solid, gel, liquid or a solid/liquid mixture that is immobilised, and the mobile phase, which may be liquid or gaseous and which flows over or through the stationary phase. The choice of stationary phase is made so that the compounds to be separated have different distribution coefficients, which describes the way in which a compound distributes itself between 2 immiscible phases.
High performance liquid chromatography (HPLC) is a form of column chromatography used frequently in biochemistry and analytical chemistry. HPLC is used to separate components of a mixture based on a variety of chemical interactions between the substance being analyzed (analyte) and the chromatography column.
In isocratic HPLC the analyte is forced through a column of the stationary phase by a liquid at high pressure. Use of pressure gives the components less time to diffuse within the column, leading to improved resolution in the resulting chromatogram. Solvents used include any miscible combination of water or various organic liquids (the most common are methanol and acetonitrile). Water may contain buffers or salts to assist in the separation of the analyte components.
The internal standard is a compound that has physical properties as similar as possible to the test analytes and which chromatographs near to, but distinct from, them.
