High Performance Liquid Chromatographic Columns

High performance liquid chromatography (HPLC) is a type of liquid chromatography that uses a liquid moblie phase. The same basic principals from gas chromatography are applied to liquid chromatography. There are three basic types of liquid chromatographic columns: liquid-liquid, liquid-solid, and ion-exchange. Liquid-liquid chromatographic columns have the liquid stationary phase bonded or absorbed to the surface of the column, or packed material. liquid-liquid chromatographic columns are not as popular because they have limited stability and they are inconvenient. Partitioning occurs between the two different liquids of the mobile and stationary phases. In liquid-solid chromatographic columns the stationary phase is a solid and the analyte absorbs onto the stationary phase which separates the components of the mixture. In ion-exchange chromatographic columns the stationary phase is an ion-exchange resin and partitioning occurs with ion exchanges that occur between the analyte and stationary phase.

Usually HPLC has a guard column ahead of the analytical column to protect and extend the life of the analytical column. The guard column removes particulate matter, contaminants, and molecules that bind irreversibly to the column. The guard column has a stationary phase similar to the analytical column.

The most common HPLC columns are made from stainless steel, but they can be also made out of thick glass, polymers such as polyetherethelketone, a combination of stainless steel and glass, or a combination of stainless steel and polymers. Typical HPLC analytical columns are between 3 and 25 cm long and have a diameter of 1 to 5 mm. The columns are usually straight unlike GC columns. Particles that pack the columns have a typical diameter between 3 to 5 µm. Liquid chromatographic columns will increase in efficiency when the diameter of the packed particles inside the column decreases.