Fast Analytical Separations with High-Pressure Liquid Chromatography

High-pressure liquid chromatography (HPLC) has established itself as a critically important analytical method for many research and commercial laboratories. Employers expect today's chemistry graduates to have a working knowledge of liquid chromatography techniques including HPLC. In addition to HPLC, it is becoming more important to educate students about newer separation technologies such as ultra-high-pressure liquid chromatography (UHPLC). Unfortunately, these systems cost significantly more than traditional HPLC instruments, preventing their broad utilization in instrumental teaching laboratories. We have developed an ultra-fast isocratic separation method using traditional HPLC instrumentation that separates five compounds in 1 min. When utilized as a teaching activity, this ultra-fast separation allows students to develop an analytical method, generate standard calibration curves, and analyze unknown samples in a single teaching laboratory period. Furthermore, chromatograms obtained with this new method are similar to UHPLC, allowing students to experience separations similar to those obtained on newer UHPLC systems using a traditional HPLC.