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Silica Gel Column Chromatography
Silica Gel Column Chromatography Description
Column chromatography is the ideal method of chromatography for purification and separation of both solid and liquids. It is a solid or liquid technique in which the stationary phase is solid adsorbents like silica gel and activated alumina powder and mobile phase is a liquid. In solvent chromatography, the selection of solvent is based on both adsorbent and solvent. The principal of active compound separation is depends on the activity of adsorbents and polarity of the solvent. If the polarity of solvent is very low and activity of adsorbent is strong and high then result of separation of compound is good. On the other hand, if the polarity of solvent is very high and the activity of adsorbents is high then it gives poor results of compound separation. It means purification and isolation of compounds are not 100% pure.
In column chromatography methods, normal phase is just opposite of reverse phase chromatography. In normal phase chromatography, stationary phase is polar and mobile phase is non-polar. Stationary phase is use polar adsorbents like silica gel and activated alumina and mobile phase is use organic and non-organic solvents like heptane, hexane, chloroform, methanol, etc.
|Size in Mesh||30-70||60-120||60-200||70-230||100-200||230-400|
|Unit in microns (µm)||210-500||125-250||74-250||63-210||74-149||40-63|
|Ph (10% aqueous solution)||7+/-0.5||7+/-0.5||7+/-0.5||7+/-0.5||7+/-0.5||7+/-0.5|
|Assay as SiO2||Min 97%||Min 97%||97.98%||97-98.3%||97-98.3%||97-99.5%|
Particle size above+
|Pore diameter-A||40-55 A||40-60 A||40-60 A||40-60 A||40-60 A||55-60 A|
|Surface Area [m2/gm]||350-450||350-450||350-450||350-550||350-500||450-650|
|LOI Loss on Ignition||<9.5%||<9.0%||<8.5%||<8.5%||<8.5%||<8.5%|
|Moisture Content %||<7.00||<6.00||<5.00||<5.00||<6.00||<5.00|
Adsorbent particle size affects how the solvent flows through the column. Finer particles (higher mesh values) are used for flash chromatography; coarser particles (lower mesh values) are used for gravity chromatography. For example, silica gel 32-63 microns and 60-210 microns is used for gravity columns and 40-63 microns for flash columns.
The process of column chromatography is the oldest and most common technique for the separation of complex mixture packed in column. Chromatography is a technology by which a mixture of chemicals are separated by its components between two phases like stationary phase which is remain fixed in placed using two adsorbents such as silica gel and activated alumina, while as mobile phase is another methods which is slowly movable and flows down though the column by either gravitational forces or external pressure into the column. Stationery phase may be solid or liquid and mobile phase is always in solid liquid foam use different solvents.
Different Methods of Chromatography:
The collectively term of chromatography is may be analytical or preparative. The starting phase of chromatography is analytical chromatography with little amount of silica gel mesh 60-120 size by using analytical column packaging, to analysis how many percentage of mixture is purify. Analytical chromatography is a simple method of chromatography with faster and cost effective separation. In analytical chemistry development, techniques for solving chemical subtracts by using thin layer plates coated silica gel on glass plate. This technique becomes standard analytical tools in pharmaceutical laboratories.
Focus on the latest chromatography technologies such as preparative and process chromatography to optimize the current and standard opportunities to optimize chromatography process in proper way. After preparative process, improving impurity profile and speed of separation use semi-preparative chromatography.
Process chromatography technology is utilized on a large scale with dynamic binding capacity. We are developing large scale process chromatography for improving purity in various biologics such as proteins, virus, vitamins, hormones, anti-bodies. In process chromatography use stationery phase adsorbents like silica gel and activated alumina are used in bulk quantity. Silica gel 230-400 and 400-800 mesh sizes are used in process chromatography to get more purity of active compounds.
The rapid foam of preparative chromatography is “flash chromatography”. To run flash column chromatography, use silica gel 200-400 and 230-400 mesh practical size in column and solvent loaded in to the column with medium air pressure of solvents which is optimized particular rapid separation and purification of targeted molecules. In flash column 40-63 microns are used for cutting-edge applications such as drug discovery, proteins and vitamins purification, chemical research and also use in other pharmaceutical research applications.
Gravity chromatography is a manual process of require continuously observed. Solvent allowed to move down the column by gravitational forced and the flow rate can be manually controlled. The practical size use of silica gel is 70-230 mesh and 63-210 microns.
Thin Layer Chromatography:
In thin later chromatography, the solid adsorbent like silica gel is coated onto a thin layer plate (about 0.25mm thick), in this case some amount of binder silica gel grades such as silica gel g (with binder), silica gel gf254, silica gel 60g,silica gel 60g, silica gel 60 gf 254, silica gel h (without binder), silica gel 60h, silica gel f254, silica gel hf254, silica gel 60h, silica gel 60hf, silica gel 60 f254 are mixed with adsorbent. In TLC grades silica gel h (without binder) is more spotlessness and high isolate grade then silica gel g (with binder). Thin layer chromatography (TLC) is simplest, easy and quick results given to the chemist as how many compounds are purify from the mixture. TLC is used in organic chemistry, a method of chemistry to identifying compounds.
Chromatography Adsorption Types:
Silica Gel (SiO2) and Alumina (Al2O3) are two adsorbents commonly used by the organic chemist for column chromatography. These adsorbents are available in different grade "mesh" size, as maintained by a number on the bottle label: “silica gel 60” "silica gel 30-70" "silica gel 60-120" "silica gel 60-200" "silica gel 70-230" "silica gel 100-200" "silica gel 200-400" "silica gel 230-400" or "silica gel 400-800" is a couple examples. This number represents to the mesh of the sieve used to size the silica, particularly the variety of gaps in the mesh or sieve through which the crude silica particle mixture is passed in the manufacturing process.