Systematic Identification of Chemical Components and Analysis of Major Constituents of Verbena officinalis L. Based on UHPLC–Q–Exactive–Orbitrap MS Combined with Feature-Based Molecular Networking and SIRIUS Strategy

Verbena officinalis L., a classic medicinal plant of the Verbenaceae family with wide clinical applications, contains alkaloids and steroids with potent anti-inflammatory and anti-psoriatic activities. However, its whole-plant chemical composition and pharmacological material basis have not been systematically elucidated. Here, UHPLC–Q–Exactive–Orbitrap MS combined with FBMN and SIRIUS software was applied for comprehensive component analysis. A total of 126 constituents were annotated following MSI standards: 15 were unambiguously identified as MSI Level 1 using reference standards, 80 were tentatively assigned to MSI Level 2 via literature MS/MS data and GNPS spectral matching, and the remaining 31 were annotated as MSI Level 3 by in silico prediction with SIRIUS. Among them, 74 compounds were detected in this plant for the first time, and 15 were preliminarily regarded as putative novel candidate constituents. This integrated method shows better isomer resolution than traditional GNPS workflows, greatly improving the efficiency and accuracy of chemical profiling of medicinal plants.