Column Packing is one of the most important step in the chromatographic purification process. A well packed column yields higher resolution and yield.

Column Packing Efficiency can be checked by calculating:

• HETP – Height Equivalent Theoretical Plates
• Peak Asymmetry Factor (AF / As)

HETP = ( d² * L ) / (5.54* t²)

Where: t = retention value expressed as units of time, volume or measured distance on the Chromatogram
d = peak width at half height expressed in same units as “t”
L = column length (in units of cm for HETP or m for N/m)

N/m =  (5.54* t²) / ( d² * L )

Asymmetry Factor (AF / As) Calculation:

AF = b/a

Where: a is the peak width (left half) at 10% of peak height, b is the peak width (right half) at 10% of peak height, 

Check out the Column Packing Efficiency Protocol

Column Packing Guidelines – TOSOH Biosciences