An integrated 3-M workflow for accelerated annotation of natural products: flavonoids in Daemonorops draco as a case study

Efficient annotation and dereplication of metabolites, particularly those from resource-endangered plants lacking reference standards, is crucial for natural products development. Advanced techniques like high resolution mass spectrometry (LC-HRMS) have significantly enhanced metabolite characterization. However, challenges such as redundant spectral data, limited reference databases, and inferior dereplication capacity hinder its broad applicability. In this study, we propose an integrated annotation strategy utilizing various computational tools, including mass defect filters (MDF), molecular fingerprints, and molecular networks (3-M strategy). We demonstrate this approach using Daemonorops draco (D. draco), a renowned yet resource-endangered natural product rich in functional flavonoids. By applying pre-defined flavonoids MDF windows, the MS