A tale of two samples: Understanding the purification workflow from different perspectives (Part 1)

Reversed-phase liquid chromatography (RPLC) is often associated with quantitative or analytical-scale analysis, but it is also a useful preparative-scale tool for purification. For any purification project, there are three possible objectives: purity, yield, and throughput. These objectives are related and are commonly represented as a triangle. Two of these targets can be achieved at the expense of the third. Prioritizing the objectives of the purification project before developing the method can help clarify the path forward. Most projects center around either bulk purification (the purification of one sample over multiple injections) or high-throughput purification (the purification of small amounts of many different samples). Bulk purifications prioritize purity and yield at the expense of throughput, whereas high-throughput purifications prioritize purity and throughput at the expense of yield. This application note by Agilent Technologies is part of a two-part series that examines the workflow for each project type and how prioritization influences the preparative workflow. Part 1 focuses on the bulk purification workflow and Part 2 gives an overview of the high‑throughput purification workflow.