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 Calculation: 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 References TOSOH Biosciences - Technical Resources GE Lifescience - Technical Resources Pall Lifesciences - Technical Resources Got something to say about this post? Leave a comment...your comments are valuable for improving the posts.