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After using an HPLC (C18) Column for several times, now it doesn't do what it has to do, when ever we want to do a research by a substance, the machine shows a every high pressure on the column and the process would be stopped on its own and won't work, we have washed is with methanol, water acetonitrile but non of them have been effective till now, what are we ignoring about this? What should we do to decrease the pressure of the column?
Available pump heads Max. flow rate in ml/min is 10
Max back pressure in bar 400
Recommended particle size is 5 μm
I have done what my friend recommended here, but none beard fruit, could the problem be something other than the column?x