HPLC

OVERVIEW

High-performance liquid chromatography (sometimes referred to as high-pressure liquid chromatography), HPLC, is a chromatographic technique that can separate a mixture of compounds and is used in biochemistry and analytical chemistry to identify, quantify and purify the individual components of the mixture.

HPLC typically utilizes different types of stationary phases contained in columns, a pump that moves the mobile phase and sample components through the column, and a detector to provide a characteristic retention time for the analyte and an area count reflecting the amount of analyte passing through the detector. The detector may also provide additional information related to the analyte, (i.e. UV/Vis spectroscopic data for analyte, if so equipped). Analyte retention time varies depending on the strength of its interactions with the stationary phase, the composition and flow rate of mobile phase used, and on the column dimensions. HPLC is a form of liquid chromatography that utilizes smaller column size, smaller media inside the column, and higher mobile phase pressures compared to ordinary liquid chromatography.

With HPLC, a pump (rather than gravity) provides the higher pressure required to move the mobile phase and analyte through the densely packed column. The increased density arises from smaller particle sizes. This allows for a better separation on columns of shorter length when compared to ordinary column chromatography.