Instrumentation Services

High-Performance Liquid Chromatography (HPLC) is a technique used for column chromatography. In this technique, a cartridge or column is packed with a sorbent (stationary phase), and a liquid (mobile phase) is passed through the packed column. A dissolved sample (in a liquid) is injected into the flow path of the mobile phase, which creates a “sample band.” As the sample band passes through the HPLC column, it separates into individual analyte bands. These analyte bands are then detected, and a chromatogram is generated, where analyte bands are seen as “peaks” and quantitated. The separation principle of HPLC is based on the distribution of the sample between a mobile phase (eluent) and a stationary phase (packing material of the column). Depending on the chemical structure of the sample, the molecules are retarded while passing the stationary phase. The specific intermolecular interactions between the molecules of the sample and the packing material define their time “on-column.” Therefore, different constituents of a sample are eluted at different times, achieving the separation of the sample’s ingredients.