chromatographic techniques - applications

chromatography - definition. column chromatography, gas chromatography, high pressure liquid chromatography, thin layer chromatography , paper chromatography - definition and application

Chromatography is a biophysical technique that enables separation, identification and purification of the components of a mixture.
The separation is based on the differential partitioning between the mobile phase and the stationary phase.
Mobile phase – solvent, which move through or over the stationary phase.
Stationary phase – either is solid with high surface area or liquid coated on to a solid; stay still.


Column chromatography is a method for separating mixtures of substances in which a liquid or gaseous solution of mixture (mobile phase) is caused to flow through a tube packed with a finely divided solid (stationary phase ), may be coated with absorbent liquid or through a long capillary tube bearing a thin film of adsorbent liquid.The components of the mixture separates based on the adsorption of the solutes of the solution.

Gas chromatography is technique used to separate and purify individual components from mixture of compounds that can be vaporized. The mobile phase is a gas and the components are separated as vapor. The separation is carried out in a column.
High Performance Liquid Chromatography or high pressure liquid chromatography relies on pumps to pass a pressurized liquid solvent containing the sample mixture through a column filled with a solid absorbent material.

Thin layer chromatography is a solid liquid adsorption chromatography. The stationary phase is applied as thin layer on a solid support plate with a liquid mobile phase.

Paper chromatography is analytical method used to separate colored chemicals or substances using a paper as the stationary phase. In this, a thick filter paper comprised the support, and water drops settled in its pores made up the stationary liquid phase. It is a liquid – liquid chromatography.

1. A pplication of Chromatographic T echniques Vasugi. M [email protected]

2. Contents Definition – chromatography. Definition and application of, Column chromatography Gas chromatography High performance liquid chromatography Thin layer chromatography Paper chromatography

3. C hromatography Chromatography is a biophysical technique that enables separation, identification and purification of the components of a mixture. The separation is based on the differential partitioning between the mobile phase and the stationary phase. Mobile phase – solvent, which move through or over the stationary phase. Stationary phase – either is solid with high surface area or liquid coated on to a solid; stay still.

4. Column chromatography Column chromatography is a method for separating mixtures of substances in which a liquid or gaseous solution of mixture (mobile phase) is caused to flow through a tube packed with a finely divided solid (stationary phase ), may be coated with absorbent liquid or through a long capillary tube bearing a thin film of adsorbent liquid. The components of the mixture separates based on the adsorption of the solutes of the solution.

5. Application of column chromatography Purification process. Isolation of metabolites from biological fluids. Isolation of active constitutes. Separation of mixture of compounds. Estimation of drug in formulation or crude extract.

6. Gas chromatography Gas chromatography is technique used to separate and purify individual components from mixture of compounds that can be vaporized. The mobile phase is a gas and the components are separated as vapor . The separation is carried out in a column.

7. Application of gas chromatography In food and beverage industry, quantitative and qualitative analysis of food composition, natural products, food additives, flavour, and aroma compounds. It is also used to analyze the presence of contamination such as pesticides, fumigants, environmental pollutants, veterinary drugs, natural toxins and packaging materials. Dairy product analysis – rancidity. Detection and determine the quantity of pesticide residue in aquaculture products. Pharmaceutical industry uses GC to ensure the purity of the produced material, to analyze compounds to check for trace contaminations.

8. In forensic, GC is used to determine the circumstance of a person’s death, such as whether they ingested poison, consumed alcohol or drug in the hour prior. Manufacturing relies on GC for quality control. In bio fuels and petroleum industry, it is used to monitor for impurities Measuring air pollution

9. High performance liquid chromatography HPLC or high pressure liquid chromatography relies on pumps to pass a pressurized liquid solvent containing the sample mixture through a column filled with a solid absorbent material

10. Application of HPLC HPLC can be used in both qualitative and quantitative application, that is for both compound identification and quantification. Separation of amino acids, peptides and proteins. Separation of sugars, lipids. Determination of cafffeine content in coffee products to gaurantee purity an quality of ground coffee. It is perfomed to maintain product purity and quality control of various industrial production.

11. Water purification. It is used for research; analyzing complex mixtures, purifying chemical compounds, developing processes for synthesizing chemical compounds, isolating natural products or predicting physical properties.

12. Thin layer chromatography Thin layer chromatography is a solid liquid adsorption chromatography The stationary phase is applied as thin layer on a solid support plate with a liquid mobile phase.

13. Application of thin layer chromatography In food industry, it is used to separate and identify colors, sweetening agent, and preservatives. Detection of pesticides and insecticides in food and water. Identification of medicinal plants and their constituents. It is used in compound purification such as amino acids, active ingredients, auxiliary substances and preservatives in drug and drug preparation. Qualitative testing of medicines like anesthetics, analgesics, sedatives, hypnotics, anticonvulsant tranquilizers and steroids.

14. It is used for biochemical metabolite separation form urine, blood plasma, serum, body fluids. It helps in analysis of fibers in forensics. It is used to track the reaction progress. Composition analysis of biomolecules or synthetic preparation.

15. Paper chromatography Paper chromatography is analytical method used to separate coloured chemicals or substances using a paper as the stationary phase. In this, a thick filter paper comprised the support, and water drops settled in its pores made up the stationary liquid phase. It is a liquid – liquid chromatography.

16. Application of paper chromatography Purification of amino acids Detection of adulterants Analysis of food colors To study the process of fermentation and ripening Detect the contamination in food and drinks It is used for drug purification, plant extract purification, isolation for abscise acid Isolation of cell wall teichoic acid of gram positive bacteria. Separation of impurities from various industrial products

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