HPLC-FLD baseline drift

[iris99]

Hi,

I've been using a Agilent 1100 with a Dionex RF 2000 fluorescence detector with an ACN/H2O system and getting positive baseline drift with increasing %ACN (ex: 260nm, em: 350nm). This did not seem to happen in the past with the same procedure, and I've tried changing to fresh solvent and extending equilibration time, to no avail and the drift is repeatable between duplicate runs. Also of note, when I switched the ACN with MeOH to see if it was a solvent issue, the same spectra was observed.

I'm not sure what I can do to solve or narrow down the problem, any suggestions or comments would be much appreciated! Thanks!

[James_Ball]

Hi,

I've been using a Agilent 1100 with a Dionex RF 2000 fluorescence detector with an ACN/H2O system and getting positive baseline drift with increasing %ACN (ex: 260nm, em: 350nm). This did not seem to happen in the past with the same procedure, and I've tried changing to fresh solvent and extending equilibration time, to no avail and the drift is repeatable between duplicate runs. Also of note, when I switched the ACN with MeOH to see if it was a solvent issue, the same spectra was observed.

I'm not sure what I can do to solve or narrow down the problem, any suggestions or comments would be much appreciated! Thanks!

Have you tried a new column? I have had FLD baseline problems when contaminates are highly retained on a column and bleed off slowly over time.

[iris99]

Hi there!

Yes this is with a new column (<1 month).