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The following message was posted to: PharmPK
We observe peak tailing with our hplc column. Recently, we faced a
problem with increased pressure and we flushed the column with buffer
free mobile phase (I have listed the general conditions below). The
organic was then gradually increased and switched to 100%
acetonitrile. The column pressure came down within a few hours and
then the eluent was gradually adjusted back to the mobile phase used.
However, after running a standard curve (50ng/ml- 20ug/ml), we
observed peak tailing with our compound of interest. The prediction
of standards and quality controls is well within the range we have
observed in our previous runs.
1) Should we be concerned about this peak tailing or can we proceed
with our analysis ?
2) How can we fix this problem ? (This column is less than 2 months old)
thanks in advance
*Buffer: * 50mM ammonium phosphate, 50mM sodium citrate and sodium
azide 10ppm ( pH adjusted to 6.5)
*Mobile Phase: *Buffer *: *methanol (82 : 18)
*Column: *BDS - Hypersil C-18, 2 x 150mm HPLC column**
*Flow rate: 0*.2 ml/min
*Injection volume:* 30ul
*Normal column pressure: *58-60 Kgf**
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Did you check the guard column? You can clean that guard column in
your mobile phase and try doing the runs again.
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