High-Performance Liquid Chromatography (HPLC) is widely used in biochemistry and analytical chemistry to identify, quantify and purify the individual components of a mixture.In our research, we focused on marine-derived fungus metabolites, which are interesting sources of natural products.The raw material extracted from the fungus is always a complex mixture.How to fractionation it and get the pure compounds is critical for further structure elucidation.With its rich column types, solvent systems and easy operating program, HPLC is a powerful tool in this field.