note

Technical
Troubleshooting Backpressure
The typical backpressure for an HPLC system is on the order of 150-250 bar (2220-3700 psi). However, the exact backpressure will depend upon many factors (Table 1). The total backpressure of the HPLC system is additive from all of the components that produce resistance to the flow of mobile phase. This includes the column, filters, cells, valves, and connecting tubes. The major proportion of backpressure will be due to the analytical column with some contribution from the guard column if present. In-line filters and coulometric cells should only contribute a few bar (each) of backpressure when working at flow-rates of 1.0mL/min or below. It is always a good idea to record the backpressure daily in a log book (make sure you take into account ambient temperature if you are not using a column oven). Monitoring the changes in back pressure provides a good diagnostic tool. The causes and remedies of lowered backpressure are presented in Table 2 while those for increased backpressure are presented in Table 3.

Decreased Pressure
A decrease or total loss of pressure is straightforward to troubleshoot. Usually this is a result of the pump not delivering adequate flow or a leak somewhere in the HPLC system. First, turn off the potentials to the cell. Check that the pump is on and operating properly. Check that there is sufficient mobile phase. If using an ESA pump, make sure that the level of fluid in the wash-bottle is normal. Examine the HPLC system from the pump to the cell, checking for leaks at each connection. Purge the pump to remove any air that may be present.

Increased Pressure
The cause of an increase in back pressure is usually more difficult to identify. If the increased pressure can be linked to an "event" (e.g., new mobile phase; different sample preparation method) then this can be used to direct troubleshooting, thereby saving downtime. However, if no such event occurred then it will be necessary to troubleshoot the whole system. Remember, before starting it is important to first turn off the potentials to the cells. Furthermore, NEVER loosen any fittings prior to the column if the HPLC system is still under pressure. Make sure that the flow is stopped and the pressure decreased to zero before breaking any connection located prior to the column. There are many approaches that can be used for investigating the cause of increased pressure. A rapid diagnostic method involves replacing the column with a union to see whether the increased pressure is due to problems with the column. If the column is the cause of increased pressure then it will need to be cleaned and/or replaced. A more methodical approach is as follows - with normal flow through the system, monitor the backpressure and see whether it changes if the fitting at the outlet of the column is loosened: If the pressure drops, then the "blockage" is located in the components located after the column (i.e., pre-cell filter, cell, or drain tubing). It is most likely to be due to the in-line filter. Replace the PEEK filter element, reassemble the inline filter and re-attach the post column components. If the backpressure increases yet again - check the cell and drain tubing (in order to prevent this problem occurring again, check Table 3 for suggested remedies).

note
Technical

Troubleshooting Backpressure
If the pressure remains elevated then the "blockage" is located prior to the column. Stop the flow and let the backpressure drop to zero. Disconnect the connection at the pump side of the first in-line filter. Then add each component, one-at-atime, turning on the flow and checking backpressure after each addition. It is most likely that the increased pressure is caused by blockage of either the first (pre-injector) or second (post injector) in-line filter (in order to prevent this problem occurring again, check Table 3 for suggested remedies). However, the smaller ID tubing used after the injection valve may be blocked or has collapsed if it was over-tightened.

Preparation of clean mobile phases, particulate free samples, and the routine changing of all in-line filters will help minimize pressure problems and downtime.

Table 1. Contributors to Backpressure.

Table 2. Reasons for Decreased Backpressure.

Table 3. Reasons for Increased Backpressure.

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