Natural products are an indispensable source for drug discovery. The major challenge for exploiting this evolutionary optimized pool of potential lead structures is the fast and reliable recognition of known compounds, i.e. dereplication. This task is essential for the discovery process in high-throughput screening scenarios, since it allows the focus to be placed on novel chemical structures at an early stage. Furthermore, information on identified compounds will help to rationalize observed bioactivities. This article describes an effective, library-supported strategy for the dereplication of crude extracts and pre-fractionated samples, using an HPLC-based multidetector platform and NMR techniques, respectively.