One hundred seventy two accessions of safflower, collected in four countries were investigated for their morphological and biochemical characters in 2014. The accessions were categorized into two groups; South-Central (S-C) Asia and South-West (S-W) Asia, and each group was represented the accessions of two countries. Variation in morphological and biochemical characters was observed between two groups of accessions. The average value of seed weight and range of variability were higher in S-C Asia accessions while S-W accessions exhibited the variation in plant height, leaf length and days to flowering. The average value of oleic and total oil content were higher in S-C Asia accessions, and the values were 19.8%, and 231.4 mg.g-1, respectively, while the range of variability for total oil content was higher in S-W accessions. Plant height exhibited a significant positive correlation with days to flowering (r = 0.625**). Palmitic acid had positively correlated with stearic acid (r = 0.282**) and linoleic acid (r = 0.444**). Oleic and linoleic acid showed a strong negative correlation (r = -0.977**). The first three principal components explained 57% of the total variation. Morphological and biochemical variation exist in different groups of accessions could be useful to breeder for developing new safflower cultivars with high oil quantity and quality.