Chromatography Forum

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

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.

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

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.