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how peak can be sharp in hplc when it is come flat
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Answer Posted / vivek k
Sample solvent stronger than the mobile phase Dissolve
sample in mobile phase or at least reduce the strength of
the sample solvent as much as possible
Sample mass overload Reduce the amount (mass) of sample
injected.
Silanol interaction with amines • Reduce mobile phase pH to
< 3.0
• Increase mobile phase ionic strength.
25mM to 50mM recommended
• Add a competing amine to the mobile phase.
10 mM TEA is usually sufficient.
• Select a stationary phase with a lower silanol activity.
Adsorption of acids on silica • Increase salt
concentration in the mobile phase
25 mM to 50 mM is usually sufficient
• Reduce the pH of the mobile phase to < 3.0
• Add a competing organic acid.
1% acetic acid or 0.1% TFA is usually sufficient
Column void Replace the HPLC column.
Attempts to fill-in the void are seldom worth the effort.
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