This study was conducted to get beneficial information of fatty acid compositions and antioxidant resources from safflower genetic resources. The fatty acid compositions and antioxidant activities in 128 safflower germplasm collected from South Asia and Africa were evaluated using gas chromatography and spectrophotometer, respectively. The total oil contents in safflower germplasm were 15.8 ~ 32.2%. The unsaturated fatty acid and linoleic acid were 89.4 ~ 93.3%, and 15.5 ~ 80.4%, respectively. Six accessions (K184652, K184658, K184662, K184671, K185831, and K185832) from South Asia exhibited high oleic acid contents. Variation of DPPH and ABTS activities ranged from 0.8 ± 0.28 to 7.6 ± 0.06 μg ASC mg-1 and from 23.1 ± 1.72 to 134.7 ± 1.25 μg Trolox mg-1, respectively. Total polyphenol contents ranged from 5.1 ± 0.17 to 52.0 ± 1.24 μg GAE mg-1. The antioxidant activities and fatty acid compositions were not different between the South Asia and Africa germplasm, while oil contents of seeds were higher in South Asia germplasm. DPPH values had positive correlation with ABTS activity (r = 0.862**) and total polyphenol contents (r = 0.864**) whereas oleic and linoleic acid showed strong negative correlation (r = −0.998**). Principal component analysis based on oil and antioxidant traits exhibited that the first four principal components together expained 85.0% of the total variation.