| Summary: | The chemical composition of <i>Vitex agnus-castus</i> L. (<i>Verbenaceae</i> family) fruits, collected from two regions in Bulgaria (south-central and north-east Bulgaria), was investigated. The content of proteins (5.3–7.4%), carbohydrates (73.9–78.8%), fiber (47.2–49.9%), ash (2.5–3.0%), essential oils (0.5%), and vegetable oil (3.8–5.0%) were identified in the fruits. The composition of the essential oils (EOs) of <i>Vitex</i> fruits from both regions was determined; the main compounds were 1,8-cineole (16.9–18.8%), <i>α</i>-pinene (7.2–16.6%), sabinene (6.7–14.5%), and bicyclogermacrene (7.3–9.0%), but significant differences in the quantitative and qualitative composition of EOs between the regions were found. The EOs of plants from north-east Bulgaria demonstrated antimicrobial activity against the pathogenic species <i>Salmonella abony</i>, <i>Staphylococcus aureus</i>, and <i>Bacillus subtilis</i>, but the Gram-negative bacteria <i>Esсherichia coli</i> and <i>Pseudomonas aeruginosa</i> exhibited resistance to the oil. Linoleic acid predominated in vegetable oil from both regions, followed by oleic acid. <i>β</i>-sitosterol and <i>γ</i>-tocopherol were the main components in the sterol and tocopherol fraction of the lipids. Phosphatidic acids were the main components in the vegetable oil from north-east Bulgaria, while in the vegetable oil from south-central Bulgaria, all phospholipids were found in almost the same quantity. Overall, significant differences were observed in the chemical composition (proteins, carbohydrates, ash and moisture) of the fruits from the two regions of Bulgaria, as well as in the content of the main components of their essential and vegetable oils.
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