Gel Permeation Chromatography (GPC)/Size Exclusion Chromatography (SEC) with Multi-Angel Light Scattering (MALS) (DAWN-HELEOS)


Gel Permeation Chromatography (GPC)/Size Exclusion Chromatography (SEC) with Multi-Angel Light Scattering (MALS)

Guarantor:
Jana Brtníková, Ph.D.

Instrument status:
Operational Operational, 30.6.2023 10:20

Equipment placement:
CEITEC Nano - B2.19 - Chromatography lab


Gel Permeation Chromatography (GPC), also known as Size Exclusion Chromatography (SEC), is an instrumental preparative analytical technique designed for the separation of polymer and biopolymers by their molecular size. Absolute molecular weight is measured using multi-angle light scattering (MALS) detection and RI detector. NECESSARY CONDITION: Solubility of tested (bio)polymers in organic solvents or aqueous buffers.


Publications:

Photogallery

Description

Complex unique GPC/SEC system with patented Wyatt detectors (MALS, viscometer, and refractometer) and separation systems (AF4 - asymmetric flow field flow fractionation) connected to the Agilent HPLC with UV-VIS detector designed for polymer and biopolymer analysis: determination of absolute molecular weight, polydispersity index, molecular weight distribution and determination of oligomers representation (aggregates), protein interactions, etc.

Applications

  • Determination of absolute molecular weight and polydispersity index without using polymer standards, molecular weight distribution, and determination of oligomers representation (aggregates), determination of the radius of gyration, second virial coefficient, intrinsic viscosity, coefficient of Mark–Houwink equation, hydrodynamic radius, and "refractive index increment" (dn/dc).
  • Analysis of (bio)polymer samples in both organic and aqueous mobile phases, is also suitable for the unique separation of (bio)polymer molecules and colloids, precise structure-size characterization of polymers, proteins and their aggregates, polysaccharides, nanoparticles, etc., including characterization of self-assembling processes in polymer-composite systems and protein interactions.

Specifications

  • HPLC system 1260 Infinity Agilent Technologies with isocratic pump, autosampler, and UV-VIS detector
  • Columns for the analysis of oligomers and polymers in the both organic and aqueous phase
  • Wyatt detectors:
    • Wyatt DAWN HELEOS II – Multi-Angle Light Scattering (MALS) detectors with 18 angles
    • Wyatt Viscostar II – differential viscometer
    • Wyatt T-rEX – differential refractometer
  • FFF-MALS system: Wyatt Eclipse asymmetric flow field flow fractionation (AF4) for the separation and fractionation of large macromolecules, biopolymers, aggregates and colloids
  • Composition-gradient light scattering (CG-MALS) system: CG-MALS Wyatt Calypso II for quantitative characterization of macromolecules, and protein interactions
  • Measuring absolute molecular weight in range 1x103 - 1x109 g/mol
  • Determination of specific "refractive index increment" (dn / dc)

Documents

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