Using High Speed-High Resolution Size Exclusion Chromatography Separation of Polymeric Materials with Light Scattering Detection

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Jennifer Gough, Michael Jones, Damian Morrison, and John Stenson
Waters and Malvern
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The need for complex product knowledge, such as for polymers and proteins, can lead to creative analytical solutions ranging from high speed chromatographic separation to advanced mass spectrometry. The novel design of the Waters Advanced Polymer Chromatography (APC) system is used to yield high resolution and quick mobile phase changes. The use of this high speed/high resolution separation technique has seen limited pairing with offline detectors such as light scattering. However, as the advantages of APC become more widely used in research and industry, the detector options that can maintain the high speed/high resolution of the APC separation are increasing.

 In this poster note, the expansion of the Waters APC approach to the size exclusion separation is presented. Sample is diverted from the ACQUITY APC Core 1 system to a Malvern OMNISEC Reveal integrated multi-detector module for analysis of refractive index (RI), ultra violet light (UV), Differential pressure viscometry (DPV), and right angle light scattering/low angle light scattering (RALS/LALS). The entire sample flow path maintains the required system dispersion control needed for the APC experiment.  The experimental samples evaluated are a mixture of two narrow polystyrene standards in terahydrofuran, and bovine serum albumin standard in auqeous buffer solution.

Coupling the APC to the OMNISEC Reveal is easily managed with the selection of the correct tubing diameter. The impact of the OMNISEC Reveal on chromatographic band spread is minimized with reduced inter-detector tubing volume and placement of the Reveal RI at the beginning of the detector flow path. In the example studied, the benefit of adding the integrated multi-detector module to the SEC experiment offers the ability to calculate absolute molecular weight, inherent viscosity and hydrodynamic radius. These calculations can predict polymer behaviors in solutions/product matrices and bring confidence to sample quality through extended quantification.


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