Liquid Chromatography–Mass Spectrometry-Based Molecular Profiling of Vertigoheel

Vertigoheel is a multicomponent medicinal product for the treatment of vertigo and dizziness, containing <i>Anamirta cocculus</i>, <i>Conium maculatum</i>, <i>Ambra grisea</i>, and <i>Petroleum rectificatum</i>. Although clinical efficacy has been reported, the chemical composition and underlying mechanisms remain incompletely characterized. Here, we applied ultra-high-performance liquid chromatography coupled to time-of-flight mass spectrometry (UHPLC-ToF-MS) to profile extracts of each ingredient and the final formulations. Untargeted analysis revealed 68,622 molecular features, and multivariate statistics highlighted ingredient-specific metabolites. Representative markers included picrotoxinin and picrotin from <i>Anamirta cocculus</i>, coniine and N-methylconiine from <i>Conium maculatum</i>, ambrinol and ambroxide from <i>Ambra grisea</i>, and santalyl phenylacetate and mercaptostearic acid from <i>Petroleum rectificatum</i>. Two compounds per ingredient were further quantified by targeted UHPLC-MS/MS, confirming their presence in drops and tablets at nanogram-per-dose levels with moderate variability across batches. These findings demonstrate that Vertigoheel retains characteristic constituents from its natural sources in all tested formulations. The established protocol enabled absolute quantification of neuroactive molecules such as picrotoxinin and coniine with minimal work-up. This molecular characterization provides new insight into Vertigoheel's composition and supports further investigation of its mechanism of action using network pharmacology approaches.