by Chromatography is a versatile analytical technique used to separate and analyze complex mixtures of substances. There are several types of chromatography, each based on different principles of separation.
- Thin Layer Chromatography (TLC):
- Principle: In TLC, a stationary phase (usually a thin layer of adsorbent material on a plate) and a mobile phase (liquid solvent) are used. The sample is applied as a spot at one end, and capillary action carries the solvent up the plate. Compounds move at different rates based on their affinity for the stationary phase.
- Uses: TLC is often used in qualitative analysis and for checking the purity of compounds.
- Gas Chromatography (GC):
- Principle: GC uses a gas as the mobile phase and a stationary phase (often a coated column or a packed column). The sample is vaporized and carried through the column by an inert gas. Compounds are separated based on differences in their partitioning between the mobile and stationary phases.
- Uses: GC is widely used in the analysis of volatile organic compounds and is especially effective for analyzing mixtures of gases or compounds that can be vaporized.
- High-Performance Liquid Chromatography (HPLC):
- Principle: HPLC uses a liquid mobile phase (typically pumped through a column) and a stationary phase inside the column. Compounds are separated based on differences in their affinity for the stationary phase.
- Uses: HPLC is one of the most versatile and widely used forms of chromatography, employed in pharmaceuticals, food analysis, environmental testing, and many other fields.
- Ion Exchange Chromatography:
- Principle: This method separates ions based on their charge. A stationary phase with charged groups binds and releases ions as the mobile phase flows through. Oppositely charged ions are attracted to the stationary phase.
- Uses: Ion exchange chromatography is used for separating and purifying proteins, nucleic acids, and other charged biomolecules.
- Size Exclusion Chromatography (SEC):
- Principle: SEC separates molecules based on their size. Larger molecules pass through the column more quickly because they don’t enter the pores of the stationary phase.
- Uses: It is commonly used in the purification and analysis of biomolecules like proteins, DNA, and polymers.
- Affinity Chromatography:
- Principle: Affinity chromatography uses a specific interaction between a biomolecule of interest and a ligand attached to the stationary phase. The biomolecule binds to the ligand while other components pass through.
- Uses: It is widely used for purifying specific proteins or other biomolecules with known binding partners.
- Reverse Phase Chromatography:
- Principle: Reverse phase chromatography uses a nonpolar stationary phase and a polar mobile phase. Compounds are separated based on their hydrophobicity.
- Uses: It is commonly used in HPLC for separating nonpolar or slightly polar compounds.
- Paper Chromatography:
- Principle: Similar to TLC, but the stationary phase is paper. The sample is applied as a spot on the paper, and the solvent is drawn up through capillary action.
- Uses: Paper chromatography is often used in educational settings and for quick, qualitative separations.
Advisory Note: Article shared based on knowledge available on internet and for the Knowledge purpose only. Please contact Professional/Advisor/Doctor for treatment/Consultation.
Articles on intactone.com are general information, and are not intended to substitute for Professional Advice. The information is “AS IS“, “WITH ALL FAULTS“. User assumes all risk of Use, Damage, or Injury. You agree that we have no liability for any damages.
