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This study evaluated the phytochemical characterization of Bixa orellana (BO extract) unsaponifiable extract and resulting fractions (F fraction - FF, Geranyl fraction - GF and R fraction- RF) obtained as by-products of an industrial process investigating in vitro antiproliferative activities in human tumoral cells. The main compounds identified by GC-MS for BO extract were Geranylgeraniol (61.51%); for FF: Geranylgeraniol (70.23%); for GF: Geranylgeraniol (78.92%) and for RF: β -cubebene (27.75%). Quantifications of geranylgeraniol by GC-FID presented the percentage content: BO 27.52%; FF 38.52%; GF 51.44% and RF 1.81%. BO extract showed a significant antiproliferative activity, with GI 50 up to 4 µg/mL. All fractions had a remarkably similar antiproliferative activity profile (GI 50 27-47 µg/mL). Data reported herein showed an important cytostatic effect for BO extract, nevertheless this activity is not attributed exclusively to geranylgeraniol. In conclusion, this by-product becomes of great value, being a potential candidate for development of new anti-tumor ingredients.