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Essay On Column Chromatography

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known as cation-exchange or anion-exchange chromatography, depending on whether the solutes to be exchanged are positively or negatively charged. Size Exclusion Chromatography: Here the molecules are separated according to their molecular weight and it is suitable for molecules having molecular weight of 2000 Daltons or more. Largest molecules are eluted first and the smallest molecules last. Affinity Chromatography: Here the stationary phase contains specific groups of molecules which can absorb the sample if stearic and charge related conditions are satisfied d. This technique is used to isolate prooteins, enzymes as well as antibodies from m mixtures. Partition Chromatography: Here the stationary phase is a thin liquid film either adsorbed or …show more content…

Sample injection system This allows introduction of the analyte mixture into the stream of the mobile phase before it enters the column. Sample injector is made by means of a syringe and self-sealing septum of silicone, neoprene, or Teflon. Modern injectors are auto samplers, which allow programmed injections of different volumes of samples that are withdrawn from the vials in the auto sampler tray. 5. Column It is heart of HPLC system made up of heavy-walled glass tubing or precision-bore stainless steel. The most common type of packing for HPLC columns is finitely divided Silica gel, Alumina and Celite. A second type of packing material is called pellicular, consists of small beads coated with layer of a porous material such as silica gel, alumina or ion exchange resin. 6. Detector This detector is used for measurement of specific physical and chemical properties of the column effluent. The most common detector used in pharmaceutical analysis is UV, which allows monitoring and continuous measurement of the UV absorbance at a selected wavelength. Appearance of the analyte in the detector flow-cell causes the change of the absorbance. If the analyte absorbs greater than the background a positive signal is

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