HPLC Column Troubleshooting Method(1)
1. High column pressure
(1) A buffer salt such as (ammonium acetate or the like) is deposited in the column;
(2) Sample contamination deposition. For this case, use 40~50°C pure water first, and rinse the column at a low speed. After the column pressure is gradually decreased, the flow rate is increased accordingly. After the column pressure is greatly reduced, rinse with normal temperature pure water, then use pure methanol.
Rinse the column for 30 minutes; for the second case, contaminate the C18 column by deposition of the sample, and rinse the column back with pure water, replace with methanol, then rinse the column with methanol + isopropanol (4+6). Rinse with methanol, rinse with pure water, and finally rinse with methanol for more than 30 minutes.
Cause There is a foreign matter between the gem ball and the valve seat with air or check valve in the system so that the two cannot be sealed. Pay attention to the amount of mobile phase in the treatment work, ensure that the stainless steel filter sinks into the bottom of the reservoir, avoid inhaling air, and the mobile phase should be fully degassed.
If there is a foreign object between the check valve and the valve seat, remove the check valve and put it into the beaker containing acetone for ultrasonic cleaning. Liquid chromatography column troubleshooting method.