How to remove peak tailing |

How to remove peak tailing in HPLC| #peak_tail_in_hplc |
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Peak tailing is the most common chromatographic peak shape distortion. We want to address how to go about fixing these distortions but first, let's understand what causes peak tailing.
Peak tailing occurs when the peak asymmetry factor (As) is greater than 1.2 - although peaks with As greater than 1.5 are acceptable for many assays.The primary cause of peak tailing is the occurrence of more than one mechanism of analyte retention. In reversed-phase separations, analyte retention is usually achieved through nonspecific hydrophobic interactions with the stationary phase. However, polar interactions with any ionised residual silanol groups residing on the silica support surface are also common. Compounds possessing amine and other basic functional groups interact strongly with such ionised silanol groups, producing tailing peaks. This is illustrated in the figure shown below at a mobile phase pHSecondary analyte interactions, with ionised silanols on the silica surface, give rise to peak tailing. These interactions need to be minimised to achieve acceptable peak shapes.
How to avoid peak tailing
There are a few methods that can be used to avoid peak tailing:
Operate at a lower pH
Use a highly deactivated column
Consider the possibility of mass overload
Consider the possibility of column bed deformation
Work at high pH when analyzing basic compounds
Use a sample clean-up procedure
If you need assistance choosing the right column, then our HPLC column selection guide will be of help.
Operate at a lower pH
As silanol groups are acidic, secondary interactions can be minimised by performing the chromatographic separation at a lower pH - thereby ensuring the full protonation of such ionisable residual silanol groups.

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