ABSTRACT
This chapter provides a brief overview of practical aspects of mobile phase preparation and column maintenance, and representing a compendium of general. Several liters of a mobile phase may pass through a column in daily use and any impurities in these mobile phases may have an adverse effect on column performance unless care is taken to minimize their presence. Almost without exception, mobile phases employed for peptide and protein separations in high-performance liquid chromatography (HPLC) are aqueous-based, with a greater proportion of water passing through a given column than any other solvent. All non-aqueous solvents commonly employed in HPLC are available in a highly pure form. If one has taken the trouble to purchase high-quality water or other solvents, or has purified the solvents on-site, it would obviously be unfortunate to then contaminate them through the use of dirty glassware. As a general guideline, any additives to a mobile phase solvent should be HPLC-grade.
