Investigation of the main active ingredients of Vitex rotundifolia L.f and assessment of their contribution to the activity of this species

• A validated HPLC method for determining active compounds in V. rotundifolia extracts. • The anti-inflammatory, analgesic, anticancer, antioxidant, and antibacterial effects of the 70 % ethanol extract of V. rotundifolia twigs and leaves. • Essential oil components of V. rotundifolia twigs and leaves and its antibacterial activity. Quality control of medicinal materials before research and use is essential. Vitex rotundifolia L.f. (also called Manjingzi in traditional Chinese medicine) is a medicinal plant with high medicinal value and uses, which is also prioritized for development in Vietnam. This study aimed to develop a validated high-performance liquid chromatography (HPLC) method for the simultaneous quantification of major compounds in V. rotundifolia for preparing the potential extract and evaluate the pharmacological effects of the V. rotundifolia extract and essential oils. HPLC method was applied for the determination of two major active compounds in V. rotundifolia , agnuside and casticin, which were determined according to the guidelines of the International Conference on Harmonization (ICH) and Association of Official Analytical Chemists (AOAC). The constituents of V. rotundifolia essential oils were also identified by gas chromatography-mass spectrometry (GC–MS). Pharmacological effects were evaluate via in vitro and in vivo experiments. The validated method demonstrated acceptable precision and was used to analyze the concentrations of agnuside and casticin in different extracts from different collection conditions of V. rotundifolia , the 70 % ethanol extract of twig and leaf V. rotundifolia collected in Ninh Hai district at noon and shade-dried was selected as a potential extract. Interestingly, the pharmacological effects of the potential extract containing agnuside and casticin were demonstrated including anti-inflammatory, analgesic, anticancer, antioxidant, and antibacterial activities. Furthermore, V. rotundifolia essential oils contain major constituents of α-pinene, sabinene, β-pinene, eucalyptol, and β-terpinyl acetate with confirmed antibacterial activity. This study has evaluated relatively comprehensively Vitex rotundifolia L.f. from chemical composition, quantitative method to biological effects, showing that this is a potential medicinal plant that can be developed in the near future.