Unveiling the dark matter of the metabolome: A narrative review of bioinformatics tools for LC-HRMS-based compound annotation.

Compound annotation, including the unveiling of dark matter in the metabolomics study represents a pivotal undertaking within the metabolomics field, serving as the linchpin for unraveling the identities and attributes of chemical entities. This narrative review examines the evolution of widely adopted compound annotation tools tailored for liquid chromatography-mass spectrometry (LC-MS) data analysis over the past two decades, which has been characterized by a transition from library-based search methodologies to advanced high-throughput approaches. Furthermore, emerging tools originating from both LC and MS domains were summarized. The synergistic partnership between quantitative structure-retention relationship (QSRR) models and machine learning (ML) techniques is explored, encompassing both conventional methodologies and advanced convolutional neural networks (CNNs). This collaborative framework has played a pivotal role in the precise prediction of retention times. Additionally, the enhanced applicability and extensibility of retention order prediction are emphasized, particularly under the constraints of experimental configurations. Within the domain of mass spectra-based annotation, the foundational task of mapping compound structures to mass spectra is examined-traditionally accomplished by aligning experimental data with established standards and libraries. Recent advancements highlight emerging tools that adopt multi-tiered mapping strategies, such as molecular networks and fragmentation trees, or incorporate machine learning to capture complex mapping patterns. This comprehensive examination underscores the pivotal role of compound annotation tools in advancing our understanding of complex LC-MS data matrix to further assist the annotation of dark matter in metabolome.