precipitation of phosphates in SEC-HPLC column
Posted: Wed Jun 12, 2013 6:10 pm
Hi all,
I just started in HPLC-SEC and I need your help.
I have run my analysis on an Agilent Bio SEC-3 column (mobile phase: 150mM phosphate buffer pH7), diconnected the column in order to check the detector and closed it provisory at both ends. it stayed like this for 2 days. precipitations were formed and I tried to redissolve the salts with the HPLC-grade water at a very low flow rate 0.005 ml/min. pressure was very high 2500-2700 psi (limit 3500 psi). I swifted to the mobile phase and after 20-30 min the pressure dropped at 10 psi and I could see the mobile phase exiting the column.
is this sudden drop of pressure normal?could the column be already destroyed? for the moment I have problems with the detector and I cannot check it.
thank you
I just started in HPLC-SEC and I need your help.
I have run my analysis on an Agilent Bio SEC-3 column (mobile phase: 150mM phosphate buffer pH7), diconnected the column in order to check the detector and closed it provisory at both ends. it stayed like this for 2 days. precipitations were formed and I tried to redissolve the salts with the HPLC-grade water at a very low flow rate 0.005 ml/min. pressure was very high 2500-2700 psi (limit 3500 psi). I swifted to the mobile phase and after 20-30 min the pressure dropped at 10 psi and I could see the mobile phase exiting the column.
is this sudden drop of pressure normal?could the column be already destroyed? for the moment I have problems with the detector and I cannot check it.
thank you