The free spreadsheet-based program HPLC Teaching Assistant was developed for effective and innovative learning and teaching of liquid chromatography. This software allows teachers to illustrate the basic principles of high performance liquid chromatography (HPLC) using virtual chromatograms (simulated chromatograms) obtained under various analytical conditions. In the first installment of this series, we demonstrate the possibilities offered by this spreadsheet to illustrate the concept of chromatographic resolution, including the impact of retention, selectivity, and efficiency; understand the plate height (van Deemter) equation and kinetic performance in HPLC; recognize the importance of analyte lipophilicity (log P) on retention and selectivity in reversed-phase HPLC mode; and manipulate or adapt reversed-phase HPLC retention, taking into account the acido-basic properties (pK a) of compounds and the mobile-phase pH.