Column chromatography is the ideal method of chromatography for purification and separation. It is a technique in which the stationary phase is solid adsorbents like silica gel and activated alumina powder and the mobile phase is a liquid. The principle of active compound separation depends on the activity of adsorbents and polarity of the solvent. If the polarity of the solvent is very low and the activity of the adsorbent is strong and high, then result of separation of compound is good. On the other hand, if the polarity of the 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.
The process of column chromatography is the oldest and the most common technique f or the separation of complex mixtures packed in a 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 method which is slowly movable and flows down through the column by either gravitational forces or external pressure into the column. The Stationery phase may be solid or liquid and the mobile phase is always in solid liquid foam use different solvents.
Analytical 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.
Preparative Chromatography: 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: 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.
Gravity Chromatography: 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.
|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|