HPLC and GPC chromatography
High pressure liquid chromatography ( HPLC) is a technique for separating, identifying and quantifying compounds. The sample to be analysed is dissolved in a liquid (mobile) phase which is pumped through a column packed with particles (stationary) phase. HPLC employs a high pressure pump to move the mobile phase through the packed column. The sample (analyte) to be analysed is injected into the stream of the mobile phase. As the analyte traverses the length of the column it is slowed by specific chemical or physical interactions with the stationary phase. When the analyte emerges from the column it passes through a detector which shows a peak with a retention time characteristic for that analyte. There are a range of detectors to suit the compounds being measured and some provide additional information such as UV spectra and chiral properties.
Click here to contact usTraditional HPLC is performed with the stationary phase coated on the outside of small spherical silica particles usually around 5 μm diameter. Stationary phases with smaller particles (<2 μm) provide more surface area with significant increases in resolution, speed, and sensitivity. However the smaller particles require much higher pressure pumps, up to15,000 psi and rapid response detectors.
Gel permeation chromatography (GPC) also known as size exclusion chromatography, separates particles on the basis of size. It is used for measuring the molecular weight of large molecules such as proteins or polymers. It is also useful for determining the tertiary structure and quaternary structure of purified proteins.
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