Calculate ligand density of a bonded phase of an HPLC column - How To
April 14, 2020
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Date: 14-APRIL2020   Last Updated: 11-OCTOBER-2025

Calculating Ligand Density for Cogent Columns

Ligand density is a key parameter that reflects how much bonded phase is present on the surface of the silica in an HPLC column. For Cogent columns, we calculate ligand density using the Berendsen-de Galan equation:

 
 
α=106×%C(102×MWcarbon×nC%C×MWligand)×SBET\alpha = \frac{106 \times \%C}{(102 \times MW_{\text{carbon}} \times n_C - \%C \times MW_{\text{ligand}}) \times S_{\text{BET}}}
α = ( 102 × M W carbon × n C % C × M W ligand ) × S BET 106 × % C

Where:α = Ligand density (µmol/m²)

  • %C = Percent carbon content of the bonded phase
  • MW₍carbon₎ = Molecular weight of carbon (12.01 g/mol)
  • nC = Number of carbon atoms in the bonded ligand
  • MW₍ligand₎ = Molecular weight of the bonded ligand (g/mol)
  • SBET = Specific surface area of the silica (m²/g)

This equation accounts for the carbon contribution from the ligand and the silica surface area to estimate how densely the ligand is bonded.

Example:
Using this method, the Cogent UDC-Cholesterol™ stationary phase has a calculated ligand density of approximately 1.5 µmol/m², indicating a well-covered and stable bonded phase.

 

 

 

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