Handroanthus chrysanthus (Synonym Tabebuia chrysantha) belong to the Bignoniaceae family and it is distributed in Central America and parts of South America. This specie is appreciated by its ornamental beauty, by the hardness of its wood and because of its broad uses in traditional medicine such as antifungal, antitumoral and anti-inflammatory. These activities are similar to those found in other species belonging to the Biggnoniaceae family such as Tabebuia impetiginosa, Tabebuia rosea and Tabebuia aurea, among others. In the present study we assessed the in vitro anti-inflammatory activity of the nhexane, chloroform, ethyl acetate, butanol and water extracts obtained from the flowers of Tabebuia chrysantha. Cell viability assays were performed to evaluate the cytotoxic effects of the extracts at three different concentrations (2 μg/mL, 1 μg/mL and 0.5 μg/mL). The concentration of 2 μg / mL exhibits cell viability percentages higher to 80% and therefore this concentration was used to stimulate murine macrophages (RAW264.7) with bacterial lipopolysaccharide (LPS) in the presence of plant extracts in order to determine the production of prostaglandin E2 (PGE2), tumor necrosis factor alpha (TNF-α), Interleukin-1 beta (IL-1β), nitric oxide (NO), and Interleukin-10 (IL-10). The n-hexane extract showed in vitro anti-inflammatory effect, inhibiting the production of PGE2, TNF-α and NO. Also, this extract induced the production of the regulatory cytokine IL-10, becoming the most promising extract evaluated. The fractionation of the n-hexane extract by column chromagraphy did generate 14 fractions (F1 – F14) and based on the chromatographic profile, the F9 fraction was submitted to a preparative chromatographic separation. After this procedure, nine fractions were obtained (F9-A – F9-I) and evaluated using GC-MS. This is the first report of the in vitro anti-inflammatory activity of the extract obtained from the flowers of Tabebuia chrysantha.