ACE C18 PFP Robustness

[JamieAlison]

Anyone out there using the 50 x 2.1 mm, 2 um particle size ACE C18 PFP column? We are using the column to analyze for CoA species and have found that the column provides great resolution. However, we are finding there to be robustness challenges with the column. After 200 injections we are seeing a tremendous loss of resolution. We are not exceeding either the recommended pH range, nor are we running the column at pressures higher than those recommended either.

We have contacted the manufacturer, who told us, after looking at the chromatography, that we probably have something in our matrix that is altering the column chemistry, leading the the resolution and poor peak shape issues. The culprit is probably protein. We are aware that there are ways to remove protein prior to analysis (we are analyzing enzyme assay samples) but are trying to run a high throughput screen to look for potential hits. I have had a hard time finding any troubleshooting information on the column online, so I was hoping that someone in the forum has experience with the column for similar analyses. Any help would be appreciated.

Jamie Alison

[Gerhard Kratz]

High throughput or long column life with sample preparation - that's the question.
200 injections with that kind of column dimension is not so bad.
Have a look at LCGC column watch. There is an article about PFP modified C18 columns. That may help.