Chromatography Forum

Dear all,

while cleaning my column and the system pre-analysis, I have chosen a created method for another analyte and the wrong mobile phase passed through my column for cca 10-15 sec (flow= 0.2 mL/min).

I stopped the flow as soon as I saw what I did, put the right mobile phases and proceeded with the cleaning and conditioning process ( after the cleaning, I leave the system to stabilize for 10 minutes with the method Im going to be using). When that was over, the column pressure was slightly higher than in the last analysis of the same analyte (instead of 181, it was 189 bar).

Is it possible that I have ruined the column it if the pressure is just slightly higher?

Run system suitability. Successful system suitability demonstrates that the system (which includes the column) is suitable.

A lot depends what the mobile phases (and column) were.

I personally think it's dangerous practice to have incompatible solvents on an hplc system. In theory if you're using only 2 bottles of 4, it doesn't matter what's in the other two bottles, but in practice it's too easy to select the wrong solvent accidentally for a moment at some stage, so I prefer the other bottles to contain something harmless.

The only time you would really worry is if you sent normal phase mobile phase through a reverse phase column or vice versa.

Something like a C18 column can handle a wide range of mobile phases, and if it was anything but a capillary column that little amount shouldn't be a problem.

What was the column and mobile phases (correct and incorrect).