Chromatography Forum

Hello

I am facing the problemof peak tailing in one of my experiments in which i have used 2mM amm acetate: Acn (10:90) as buffer and acn:water(90:10) as diluent.

And i have used normal C18 column. And also tested with cyno as well as phenyl columns but the tailing remains with the peak. can you any body help me to solve this problem?

And i also tried with different compositions of mobilephase using methanol and added THF and so many trails i have done.

You really haven't given enough information:
- what is the nature of your analyte peak (acid, base, . . .)?
- can you be more detailed about your conditions. From your post, I surmise that your sample is dissolved in the "diluent" and that "buffer" is your chromatographic mobile phase. If so, 2mM is somewhat on the low side for buffer concentration.
- what pH is your buffer?
- what is the injection volume?
- can you be more specific about the columns you have looked at. Basic compounds in particular are much more prone to tail on older "type A" silica-based columns (regardless of bonded phase).

Dear Mass

Could u ple. inform here what tailing factor U got ?

If U can clean yr column with hot water ( about 50 deg. C.for 2 hours ) will give u some advantages .

If yr method is validated on the same mobile phase then it is a matter of cleaning of column . You may change mobile phase ACN & Water ( 90 : 10 )

If u r just trying to develop the method , ple. donot try this way .U check following first before analysing on HPLC .

1. Polarity of compound

2. Nature of compound -- acidic or Basic

3. What is the lamda maxima on UV scanning