Air was flowing for 3 days into my column..

[lsreedharan]

Hello everyone,

Im using ZOrbax eclipse XDB- C8 column to separate bile acids and fecal sterols. Unfortunately, last week i stopped working on LC-MS and I left my column jus like that. Yesterday I realized that my pumps were on and solvents were flowing at .5 mL/ min and pump program was 65% of 0.2 mM ammonium acetate in water and 35% of methanol. I was totally out of methanol and I calculated the amount of methanol in the bottle and it was out in the very second day. Only the aqueous phase was flowing from pump A all these days (5 days) and from pump B just air. What should I do now? Will my stationary phase will be completely dried? Please help me regarding this. Thanks in advance.

Thanks,
Sree

[tom jupille]

Most (but not all) pumps will not pump air (that's what "losing prime" is all about). Best advice I can give is to flush the column for a bit and then test it. If it's already dead, that won't kill it any further.

[lsreedharan]

Thanks for your advice Mr.Tom, I will flush the column and try for my luck.

[Gerhard Kratz]

I would flush the column with a low viscosity solvent, for example ACN, with a very low flow rate. But first flush the system without the column. And to avoid additional damage I would flush the column by reversed flow direction.
Good luck.