[Protein-analysis] Re: Help me! AIR BUBBLES trapped in HPLC column...

Dr Engelbert Buxbaum via proteins%40net.bio.net (by engelbert_buxbaum from hotmail.com)
Tue Jan 12 02:21:06 EST 2016

In article <mailman.309.1452553859.6180.proteins from net.bio.net>, majedbilal from gmail.com says...

> After I had completed my analysis, I switched to sequence flush overnight for two hours. 
> However I made an error in the settings and realised in the next morning the method was >
still running and I ran out of wash phase. So air was being pumped into the system and the > 

One thing you can try is to mount the column vertically and to pump medium from below. Use 
something with relatively low viscosity like acetonitrile or methanol. Afterwards measure the 
number of theoretical plates of the column to see how much resolution you have lost.

The second option is to repack the column, however, for HPLC that requires a packing kit. 
Open the column, empty the stationary phase into an vacuum flask with, say, 20% methanol and 
connect to vacuum. Once no more air bubbles come out break the vacuum and use the slurry to 
repack, following instructions that came with the kit. 

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