Chromatography Forum

Hi,
i'm using an AGILENT Prep HT Eclipse XDB-c18 (150x21.2-5µm) column for purification of several kinds of molécules.
The problem is that i got peak splitting with a ratio of 20-80 for the amount of split with every kind of product, so the sample solvent can't be the cause.
I would like to know if you can tell me a recovery procedure to fix the problem or maybe the column is "dead" :-[ ....
I use a column guard of the same stationnary phase but it seems it has not protected the column ....
I work for an R&D outsourcing laboratory in drugs research so i can't tell you what was the product i use when the problem apeared...., the eluent system is ACN-water or sometime TFA 0.1% aq. buffer.
Thanks

what are the suggestions from Agilent technical persons?

Column stability on prep columns is a non-trivial business. I suggest to look at another vendor that has solved the problem, i.e. Waters.

I am usually not that bland, but there is really a large difference in the technology between different vendors, and it is worth your while to investigate, especially considering the price of such columns.

The problem seems to be solved:
I used the column inverted, washed with Water and ACN 95-5 to 5-95 and 5 injections of 200µL of DMSO.... so for the moment, i don't have any peak splitting, just a little tailing....
Thanks for your replys.

Nicolas

Nicolas,
I’m also having the peak splitting problem. The column that I’m using is Bondapak phenyl, 3.9 x 300 mm 10 . Why did you inject DMSO five times after washing your column?
Thanks,

Provetech,
I might suggest a possible reason for the peak splitting might be partial clogging of the inlet frit on the column. I have seen this split peaks before, and lloks very much like a column void. Maybe your pressure increased slightly due to some insoluables or ??? Anyway just another thought about the splitting.

Regards,
Mark