Leakage is a common problem in HPLC analysis. To minimize leaks in your system, try to choose hplc instrument parts from the same manufacturer. If an incompatible hplc joint is installed, problems may occur during separation and repeated connections may eventually result in a leak of the joint. If replacement is required, use the appropriate fitting connector and check all connections for leaks prior to testing.
Chromatographic column frits clogging is another common PROBLEM with HPLC. To minimize this problem from the start, use a guard column. The chromatographic inline filter can filter out pollutants in the mobile phase and protect the hplc column from precipitates in the sample solution.
To clean the injector normally, disconnect and reverse the column, connect it to the hplc pump (but not to the detector), and pump the solvent at twice the standard flow rate. Approximately 5-10 column volumes of solvent should be sufficient to remove small amounts of granular material from the inlet sieve plate. Finally, a standard test mixture was used to evaluate the performance of the cleaned column.
Regular replacement of chromatographic column frit is recommended. Sometimes neither solvent flushing nor recovery procedures can restore column performance. If you have identified the column as the source of the problem and repair procedures have failed for other columns, there may be gaps in the packing or blocking of the inlet sieve plate. As a last resort, open the inlet end of the column. Note: Opening the inlet end, especially the outlet end, may permanently damage the packed bed. Consult the manufacturer’s literature before opening the column.
High performance liquid chromatography troubleshooting. Disconnect the column from the system. To prevent the filler from leaking out of the column, perform the following steps as soon as possible. Using a vise and wrench, or two wrenches, carefully remove the inlet end connector. If the frit remains in the joint, remove it by striking the joint on a hard surface. If the sieve plate is left on the column, slide it down rather than lift it up. This will help maintain the integrity of the packed bed.
Modular columns may require special tools to remove the sieve cap. Inspect old frits. Pressing the screen plate against the stainless steel tube will leave a ring around the column side edge of a properly installed screen plate. The absence of a ring may mean that the ring is too close to the end of the tube. The resulting loose connections may leak silica or act as mixing chambers. Check the packed bed. If it sags or breaks, a new column is needed. Replace the hplc frit . Replace the end connector. Tighten by hand, then use a wrench to tighten 1/4 turn.
Post time: May-28-2022
