Chromatography Forum

This may be due to the recent samples we've been getting (despite the fact sample prep involves a 0.22 um filtration), but we've been having heavy peak tailing issues on our Agilent HPLC system.

We would flush the column with mobile phase (we run an isocratic run with 0.005 N sulfuric acid on a Supelcogel C-610H 9 um 30x7.8 cm column) - reverse the column and the guard column so the flow is in the opposite direction - and it would fix the problem at first. But after even a few sample injections the tailing would appear again.

I realize the dirty samples might not be helping, but any advice to keep the tailing at bay for longer? Such as good flush solvents or guard columns to help keep the system clean enough? We just bought the column 3 months ago.

Thanks so much!

I would speculate that the guard cartridge (or possibly, but less likely the column) has a head space. When you reverse the flow, it eventually moves to the other end. If this were my problem, I would start by changing the guard cartridge.

Completely agree with Tom as a void at the head of the column is the most likely (and in fact very common) reason. These gel columns are sometimes quickly compressed, fouled and/or damaged. It is not your HPLC system's fault. Columns are consumable items, meant to be replaced. If the column is not lasting very long, then review your method to insure it follows good chromatography principles, do not backflush gel columns and of course review your sample prep procedures. Have someone with HPLC experience review your method and techniques on site to find the true cause and make suggestions or provide training to solve.