Ionisable Lipid Purification with Centrifugal Partition Chromatography Under Water-Free Conditions

This study examined the purification of ALX-24, a hydrophobic ionisable lipid, following synthesis. The starting material had a QNMR content of approximately 88%. The purification was carried out on a 34ml miniLiLi column in descending mode using a completely water-free solvent system consisting of hexane, MTBE, acetonitrile, and acetic acid. A 150 mg sample was injected, and the fractions were collected according to the UV signal at 210 nm. The main product fraction reached 99% purity by QNMR with a 70% yield. This demonstrates that CPC can purify hydrophobic ionisable lipids without introducing water into the process—an important consideration for moisture-sensitive lipid materials and downstream applications. Key result: The ionisable lipid was purified from approximately 88% to 99% purity with a 70% yield under entirely water-free conditions