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What is Chromatography?
Chromatography has been developed into a new method of separation of mixture of compounds mainly when they are available in small quantities. This method is very useful when the components of a mixture have almost the same physical and chemical properties and hence cant be separated by other usual methods of separations. The term chromatography means writing in colour (Chroma = Colour & Graphy = To write).
Types of Chromatography
Paper Chromatography
Gas Chromatography Thin Layer Chromatography
The stationary phase or adsorbent in column chromatography is a solid. The most common stationary phase for column chromatography is Silica Gel, followed by Alumina Oxide.
The mobile phase or eluent is a liquid. It is either a pure solvent or a mixture of different solvents. It can be used for molecules whose molecular weight is < 2000 g/mol
Uniform layer of Silica Gel coated onto a piece of glass, aluminium or rigid plastic plates are being used for analysis.
The mobile phase moves through the stationary phase and carries the components of the mixture with it. Different components travel at different rates. Application: To determine the number of components in a mixture, Identification of compounds, and Purity of a compound.
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In Thin Layer Chromatography fluorescent is added in stationary phase to analyze substances which are colourless. This we can not see with our naked eyes. When plate will expose to UV light then it will Glow except the spots of substances & spot will look like a dark patch.
Column
Semi Preparative: Column Inner Dia - 10, 20, 30 mm & it is done at milligram scale.
Preparative: Column Inner Dia 40, 60, 80, 80 mm & it can be done at gram Scale Pilot scale: Kg scale (1, 2, 4, 10, 12 Kg) Commercial scale: Bulk Quantity (50, 100, 200, 400 Kgs or more)
Allow the solvent to move though the column until reaches to the bottom. At this stage vacuum is not require.
Allow 56 columns value of solvent to flow through the column to make sure it is complete packed.
Drain the solvent till the solvent level is just even with the surface of the stationary phase
Types of Columns
Gravity Column Chromatography: Solvent is allowed to move down the column by gravitational forces. Flash Chromatography: Solvent is pushed down the column by positive air pressure
Application
Separation of mixture of compounds Purification process Purification of Phytochemical Isolation of metabolites i.e. Small molecules Estimation of drugs
Process Development
Purify Natural compounds To separate active component from Plant material Herbal Extraction
These adsorbents are sold in different mesh sizes such as 60-230 mesh, 100-200 mesh, 200-400 mesh & tailor made.
Adsorbent particle size affects how the solvent runs through the column. Smaller particles (higher mesh size i.e. 230-400 mesh) are used for flash chromatography & larger particles (lower mesh size i.e. 60120/60-200) are used for gravity chromatography.
Reverse Phase Chromatography It uses a non polar stationary phase and a polar mobile phase. Polar compounds elute faster than non polor compounds. When we increase polarity of mobile phase elution time will decrease. It Can reused / reproducible
Mobile phase are non polor i.e. IPA, hexane, dichloromethane, chloroform, ethyl ether, and isopropyl alcohol (IPA).
Disadvantages
Pharmaceutical Industries Bulk Drugs & API Nutraceuticals Herbal Extracts products manufacturers
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III. Novel Drug Discovery, IV. Clinical Research, V. Pilot Scale lab,
Research Laboratories
Laboratories Chemical Repackers Contract Research Laboratories
Silent Features
Manufacturing since 1973 Consistence supplies The product offered is highly active material Our products has got higher surface area
Aluminas Surface Area is less than Silica Gel i.e. 140-160 m2/gm Size Available: 100-300 mesh & 200-400 mesh