HPLC
High pressure liquid chromatography (HPLC) is a technique that uses a solid stationary phase, packed into stainless steel tube or column and a liquid mobile phase (acetonitrile, methanol, phosphate buffer etc..) to separate out components in a mixture. Chemical interactions between the stationary phase and the chemical composition of the components in mixture result in the components to travel at different speeds in the column and separate out or elute at different times from the column stationary phase.
Common LC Column Formats
Nano LC Columns
High powered sensitivity for exceedingly small samples (Column ID 50-75 μm)
Micro LC Columns
Increased sensitivity for small samples (Column ID 0.15-.05 mm)
UHPLC Columns
High speed separations of analytes (Column ID 1.0-2.1 mm)
Analytical LC Columns
General, all-purpose separations (Column ID 2.1-4.6 mm)
Semi-Prep and Preparative LC Columns
Large scale isolation and purifications (7.8-100 mm)
| Separation Mode | Description: |
|---|---|
| Reversed Phase | Used to separate hydrophobic compounds. |
| Normal Phase | Used to separate hydrophobic compounds and matrices that are retained too strongly by reversed phase. |
| HILIC | Used to polar organic compounds that are poorly retained by reversed phase. |
| Ion Exchange | Used to separate charged compounds. |
| Ion Exclusion | Used to separate organic acids, carbohydrates, sugars, starches, and oligosaccharides. |
| Chiral LC | Used to separate enantiomers. |
| Size Exclusion Chromatography (SEC) | Used to separate biomolecules and polymers. |
