ABSTRACT
This chapter presents some practices that will help improve the performance and lifetime of liquid chromatography (LC) columns used for protein and peptide analysis. Topics include sample preparation, mobile phase composition, in-line filters, guard columns, and precolumns. Troubleshooting techniques cover how to deal with blocked frits, column voids, column contamination, and bonded phase loss. The best way to assess the quality of an LC column is to validate its performance when new and continue to use the same validation protocol throughout the lifetime of the column. The best way to maximize the performance and lifetime of an LC column is to practice regular preventive maintenance. The most powerful means of preventing column problems in the LC separation of biopolymers is to take the time to prepare properly the samples to be injected. For all types of LC columns, it is important that the mobile phase components are miscible with each other and with the injection solvent.
