types of columns in High performance liquid chromatography.pptx

this presentation gives detailed information about types of columns used in HPLC

1. TYPES OF COLUMNS IN HPLC SUBMITTED BY Aiswarya S V (12900) PhD 1 st year Division of Microbiology AC 509- SPECTROSCOPY AND CHROMATOGRAPHIC TECHNIQUES

2. Normal-Phase Columns (NP-HPLC) : Stationary Phase : Polar Common Materials : Silica, cyanopropyl Applications : Separation of polar compounds, such as organic acids, carbohydrates, and natural products. Reversed-Phase Columns (RP-HPLC) : Stationary Phase : Non-polar or moderately polar Common Materials : C18 (octadecylsilane), C8 ( octylsilane ), phenyl Applications : Widely used for a broad range of organic compounds, including pharmaceuticals, peptides, and proteins. Ion-Exchange Columns (IEX-HPLC) : Stationary Phase : Charged (anionic or cationic) Common Materials : Sulfonic acid (for cation exchange), quaternary amines (for anion exchange) Applications : Separation of charged molecules like amino acids, peptides, proteins, and nucleotides.

3. 4. Affinity Columns : Stationary Phase : Ligands or antibodies specific to the target molecule Common Materials : Agarose, silica beads functionalized with ligands Applications : Purification and separation of biomolecules, such as enzymes, antibodies, and hormones. 5. Chiral Columns : Stationary Phase : Chiral selectors Common Materials : Cyclodextrins, polysaccharide derivatives Applications : Separation of enantiomers, crucial in pharmaceutical analysis. 6. Hydrophilic Interaction Chromatography (HILIC) Columns : Stationary Phase : Polar Common Materials : Silica, amino, cyano Applications : Separation of highly polar compounds, such as peptides, amino acids, and nucleotides. 7. Size-Exclusion Columns (SEC-HPLC) : Stationary Phase : Porous Common Materials : Cross-linked polystyrene, silica Applications : Separation based on molecular size, used for polymers, proteins, and polysaccharides.

4. DIFFERENCES BETWEEN NP Vs RP HPLC