Lecture

Size Exclusion Chromatography

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In course

CH-301: Analytical separation methods

Les étudiants comprennent les bases physico-chimiques des méthodes de séparation chromatographiques et électrophorétiques. ...

Description

This lecture covers the principles and applications of Size Exclusion Chromatography (SEC), also known as Gel Permeation Chromatography (GPC). SEC is used for analyzing macromolecules based on their molar masses, with the eluent typically being a single solvent. The lecture explains the interpretation of chromatographic data, the different types of columns and detectors used, as well as the stationary phases involved. It delves into the retention principles, distribution coefficients, and the factors influencing the separation process. Various detectors like refractive index, UV-visible, and light scattering are discussed, along with the importance of column temperature and sample injection volume. The lecture also touches upon the different types of gels used in SEC, the elution process, and the factors affecting resolution and selectivity.

Instructor

Christophe Roussel

Habilitation à Diriger les Recherches (HDR) in chemistry at the Ecole Normale Supérieure Paris-Saclay (Cachan, France)PhD Thesis in physical organic chemistry at the University of Bourgogne Franche-Comté (Besançon, France) / Ecole Normale Supérieure Paris-Saclay (Cachan, France)

Official source

Related concepts (29)

High-performance liquid chromatography

High-performance liquid chromatography (HPLC), formerly referred to as high-pressure liquid chromatography, is a technique in analytical chemistry used to separate, identify, and quantify each component in a mixture. It relies on pumps to pass a pressurized liquid solvent containing the sample mixture through a column filled with a solid adsorbent material. Each component in the sample interacts slightly differently with the adsorbent material, causing different flow rates for the different components and leading to the separation of the components as they flow out of the column.

Elution

In analytical and organic chemistry, elution is the process of extracting one material from another by washing with a solvent; as in washing of loaded ion-exchange resins to remove captured ions. In a liquid chromatography experiment, for example, an analyte is generally adsorbed ("bound to") an adsorbent in a liquid chromatography column. The adsorbent, a solid phase (stationary phase), is a powder which is coated onto a solid support.

Molar mass

In chemistry, the molar mass (M) of a chemical compound is defined as the ratio between the mass and the amount of substance (measured in moles) of any sample of said compound. The molar mass is a bulk, not molecular, property of a substance. The molar mass is an average of many instances of the compound, which often vary in mass due to the presence of isotopes. Most commonly, the molar mass is computed from the standard atomic weights and is thus a terrestrial average and a function of the relative abundance of the isotopes of the constituent atoms on Earth.

Macromolecule

A macromolecule is a very large molecule important to biological processes, such as a protein or nucleic acid. It is composed of thousands of covalently bonded atoms. Many macromolecules are polymers of smaller molecules called monomers. The most common macromolecules in biochemistry are biopolymers (nucleic acids, proteins, and carbohydrates) and large non-polymeric molecules such as lipids, nanogels and macrocycles. Synthetic fibers and experimental materials such as carbon nanotubes are also examples of macromolecules.

Liquid chromatography–mass spectrometry

Liquid chromatography–mass spectrometry (LC–MS) is an analytical chemistry technique that combines the physical separation capabilities of liquid chromatography (or HPLC) with the mass analysis capabilities of mass spectrometry (MS). Coupled chromatography - MS systems are popular in chemical analysis because the individual capabilities of each technique are enhanced synergistically. While liquid chromatography separates mixtures with multiple components, mass spectrometry provides spectral information that may help to identify (or confirm the suspected identity of) each separated component.

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