Aim of chromatography
• The compounds of interest has to be separated from matrix components
• The signal to noise ratio must be as high
as possible
• The time consumption of analysis must be short
Mobile Phase / Stationary Phase
Phases Involved in Chromatography
Mobile Stationar phase phase y Type of chomatography
Gas Gas Liquid Solid Liquid Solid Gas Solid Chromatography Gas Liquid Chromatography Liquid Solid Chromatography
Liquid Liquid Liquid Liquid Chromatography
Retensi dalam Kromatografi
Sampel bergerak dari fase bergerak ke fase diam (tertarik) / sorpsi , dan terjadi kebalikannya desorpsi
Chromatographic process
• The injected sample is dragged by mobile phase
along the column.
• The components of the sample distribute
between the stationary phase and mobile phase.
• If X compound has bigger affinity to stationary phase than affinity of Y compound to stationary phase, the X compound elutes later than Y from column.
• The sharp injected peaks become broader and
broader during their run.
Geometry of Chromatography System
• Column chromatography
the stationary phase is contained in a tube
• Planar chromatography
the stationary phase is configured as a thin two- dimensional sheet in paper chromatography (a sheet or a narrow strip of paper); or
in thin-layer chromatography (a thin film of solid particles bound together for mechanical strength with a binder, such as calcium sulfate and coated on a glass plate or plastic or metal sheet)
Column Chromatography
• Column chromatography & liquid chromatography (typically mean) : the stationary phase, adsorbed on a solid support to produce a packing
• Original stationary phase is water adsorbed onto solid particles (powdered sugar, powdered cellulose, or other possibilities)
• Packing is often packed into a glass tube about 20 mm in diameter and 200-300 mm in length by tamping it with a glass rod
• Sample is applied into the top of column and
allowed mobile phase to pass by gravity flow
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