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SEC Column Selection for Polysaccharide Molecular-Weight Determination (60–70 kDa)

Record: 640

Category: Column Selection
Product: SB-803
Sample: Polysaccharide

Question

A customer has the following Shodex columns available and would like to determine the molecular weight of polysaccharides using size exclusion chromatography. The expected molecular weight of the sample is approximately 60–70 kDa.

Available columns:

Which of these columns is most appropriate for this analysis?

Answer

Among the listed columns, Shodex OHpak SB-803 HQ is the most appropriate choice for molecular-weight analysis of polysaccharides in the 60–70 kDa range.

SB-803 HQ is a polymer-based aqueous SEC column specifically designed for water-soluble polymers and polysaccharides. Its effective separation range spans approximately 10 kDa to several hundred kDa, placing a 60–70 kDa polysaccharide well within its optimal operating window. This column is commonly calibrated using pullulan standards, P-82 kit, which are linear polysaccharides and provide a reliable reference for estimating polysaccharide molecular weight.

The other available columns are less suitable for this application. PROTEIN KW-803 is optimized for protein separations and molecular-weight determination of globular proteins rather than neutral polysaccharides. Asahipak GF-series columns are primarily intended for biomolecules such as proteins and nucleic acids and are not optimized for neutral or weakly interacting polysaccharides.

Based on pullulan calibration data, a 60–70 kDa polysaccharide would be expected to elute between the P-50 (Mp ≈ 47,000) and P-100 (Mp ≈ 107,000) standards, which aligns well with the separation characteristics of SB-803 HQ.

Recommended Application:

SEC-based molecular-weight determination is relative and depends on polymer conformation and calibration standards. Pullulan standards are widely used for polysaccharide analysis, but absolute molecular weights may differ for branched or structurally distinct polysaccharides.

For assistance for a more tailored column selection, eluent selection, or calibration strategy, please contact Shodex Technical Support at support@shodexhplc.com.

Additional Comments

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