how peak can be sharp in hplc when it is come flat
Answers were Sorted based on User's Feedback
Answer / sasikumar.r
1. increase the concentration of organic phase. i.e.,
increase the ratio of organic phase in mobile phase
2. Add little amount of peak modifiying agent TRIETHYLAMINE
3. just REVERSE the column
| Is This Answer Correct ? | 15 Yes | 5 No |
Answer / 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.
| Is This Answer Correct ? | 10 Yes | 0 No |
Answer / shashikumar.v
Add little amount of peak modifiying agent TRIETHYLAMINE
Add 1% acetic acid
Reduce the pH of the mobile phase to < 3.0
| Is This Answer Correct ? | 1 Yes | 0 No |
Answer / pragya
By the small addition of organic solvents or by changing the mobile phase ratio with the small change in organic modifiers.
| Is This Answer Correct ? | 1 Yes | 0 No |
Answer / anandpcp
Drug is a any chemical or a chemical substance substance
which are used for the specific purpose cure, prevention
etc...i.e, treatment of any disease or disorder for humen or
animal.
| Is This Answer Correct ? | 1 Yes | 5 No |
Answer / dhanalakshmi
You can use Peak height or Peak area to calculate the
concentration and the results are no big different. My
experience is to see which one is sharp on the chromatogram.
E.g. if a very sharp peak was found then I will use the peak
height, if a flat peak then I will use the peak area for
calculation
| Is This Answer Correct ? | 3 Yes | 15 No |
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