Monday, December 20, 2010

Peak tailing, Peak Fronting, Rounded Peaks in HPLC chromatogram

Peak tailing



What you expect                                   What you Got



Disease
Medicine
Active sites within the column
Test with standard test mixture. If OK add competing base or acid modifier
Wrong pH
Correct
Wrong column
Change
Void volume at inlet
May need repacking
Dissolve sample in mobile phase



Peak fronting

What you Expect                                What you Got    



Disease
Medicine
Column overload
See this (Retention Times are not constant)
Wrong pH
Correct
Sample solvent incompatible with mobile phase
See this (Peak Tailing)
Void volume at inlet
May need repacking
Wrong sample solvent
Dissolve sample in mobile phas





Rounded peaks




What you Expect                                  What you Got



Disease
Medicine
Detector outside linear dynamic range
Reduce sample
Gain too low
Adjust
Column overloaded
See this (retention times are not constant)
Time constants (detector, recorder) too high
Reduce
Wrong sample solvent
Dissolve sample in mobile phase


Reference:  
Prof. Dr. H.J. Chaves das Neves
Universidade Nova de Lisboa

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