Abstract Vitex negundo L. (Verbenaceae) is found in south- eastern Asia, Afghanistan, India, China, Tanzania, Madagascar, Europe, West Indies and North America as a much branched aromatic shrub. The plant parts are used to treat asthma, body pains, diarrhoea, dysentery, dyspepsia, fever, liver, skin and spleen diseases, malaria, piles, rheumatism, sinusitis, spermatorrhoea, sprains, swellings, urticaria, vomiting, weakness and whooping cough. Phytochemical investigation of a methanolic extract of the stem bark led to isolate ten chemical constituents characterized as the fatty esters n-undecanyl oleate (1), n-undecanyl stearate (2) and n-tetracosanyl capriate (6), long chain alkanes n-pentacosane (3), n-henetriacontane (4) and n-pentatriacontane (5) and triterpenoids lanostan-3-olyl 1′-(3′- eicosanyl) anthraquinone (7), lanostan-3-olyl 1′-(4′-arachidyloxy)-xanthone (8), oleanolic acid 3-O-α-D-glucopyranosyl-2ʹ-arachidate (9) and betulinic acid 3β-O-α-D-glucopyranosyl-2′-lignocerate (10). The structures of all the isolated phytoconstituents have been established on the basis of spectral data analysis and chemical reactions.