14 Types of Chromatography (Definition, Principle, Steps, Uses)

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.

  1. 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.
  2. 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.
  3. 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.
  4. 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.
  5. 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.
  6. 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.
  7. 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.
  8. 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.

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