Column chromatography is a separation technique that is used to purify compounds based on their hydrophobicity or polarity. The principle of column chromatography depends on the differential adsorption of a solute by the stationary phase (adsorbent). The mixture of complex molecules of analytes are separated based on differential partitioning among a stationary phase and a mobile phase, there are various sizes of the column are available for this technique.
In the process, the analytes to be separated are placed on top of a solid adsorbent-packed column. Then the mobile phase is loaded into the column at the top and the column is allowed to flow slowly and continuously. Components with less adsorption and affinity toward the stationary phase travel rapidly than those with higher adsorption and affinity for the stationary phase. Fast-moving components are eluted first, followed by slow-moving components.
Different types of column chromatography such as adsorption column chromatography, partition column chromatography, gel column chromatography, and ion-exchange column chromatography are used to separate the active ingredients by various methods.
Applications of column chromatography:
- Column chromatography can be used to isolate many classes of drugs and components such as glycosides, plant extracts, alkaloids, amino acids, formulations, and drugs.
- Used for separation of the mixtures of compounds.
- It is used to remove impurities or the purification process.
- In column chromatography Impurities in a compound can be separated by using the appropriate mobile phase and a stationary phase.
- It is applied to the separation of active constituents.
- From the crude extracts, formulation, active constituents, plant extract necessary constituents can be isolated by this type of chromatography.
- Column chromatography is used for the estimation of the drug in the formulation.
- From the biological fluids, metabolites can be separated using column chromatography.
- Using this method, it is possible to the estimation of the drug in the crude extract.
- For the isolation of active ingredients from plants, column chromatography is the most preferred and only separation technique in phytochemistry.
Chromatography is commonly used in chemical and life science research for a variety of purposes. The importance of chromatography techniques can be seen in their widespread use in a variety of sectors for a variety of objectives. HPLC, GC, TLC, adsorption, partition, affinity, and paper chromatography are some of the types of chromatography that have several applications in different fields.
General applications of chromatography are pharmaceutical industries, food industry, fuel industry, environmental analysis, forensic science, biotechnology, biochemical processes, and biological application, molecular biology studies, etc.
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