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What do I specify as the Void Volume in Empower when running system suitability testing with BEH SEC columns? - WKB96881

Article number: 96881

ENVIRONMENT

  • Bridged Ethylene Hybrid (BEH)
  • Size Exclusion Chromatography (SEC)
  • System Suitability
  • Calculations
  • Void Volume
  • Total Exclusion Volume

ANSWER

The Total Exclusion Volume should be specified as the "void volume".

ADDITIONAL INFORMATION

In SEC, analytes are not retained since there is no adsorption mechanism in SEC as there is in reversed-phase. 

When an analyte is too large to enter any of the pores on an SEC column, it is excluded from entering any pores and elutes before any other analytes.

Therefore, the analyte that is too large elutes in what is referred to as the "Total Exclusion Volume".

On a BEH200 SEC column, the Thyroglobulin dimer from the BEH200 SEC standard elutes in the total exclusion volume. 

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