Rapid peptide separation by supercritical fluid chromatography

Background: Supercritical fluid chromatography (SFC) is continually gaining attention in the separation sciences as demand increases for higher throughput isolations and purifications. The higher flow rates associated with SFC provide a significant decrease in analysis time and increase in sample-throughput efficiency. Peptides are of particular interest for SFC due, in part, to the rather extensive analysis time required by HPLC. Results: This work explored a wide range of peptides not only for detection, but also for separation using SFC. A separation of five peptides ranging in molecular weights from 238.2 to 1046.2 was achieved by SFC in less than 12 min, compared with 50 min using HPLC. Conclusion: This research further illustrates the ever-expanding applicability of SFC to a wider variety of compound classes. The rapid analysis time associated with SFC, as seen in this work, provides a nearly fivefold decrease in analysis time when compared with HPLC.