Carotenoids of squash play an important role in human health by acting as sources of provitamin A or as protective antioxidants. Among the 60 accessions of squash germplasm, fluorescent yellow and yellow types of flesh color got the highest count, followed by the orange, whitish yellow and greenish yellow. The redness and yellowness values of the flesh powder ranged from -2.45 to 86.09 and from 13.77 to 39.80, respectively. While the lightness and the total color difference values of flesh color varied from 67.64 to 86.09 and from 19.77 to 51.79, respectively. Colorimetric values of redness and yellowness showed positive correlation, and the correlation coefficient (r) was as high as 0.7386. The five accessions represented each flesh color types, IT195043 (orange), IT136696 (fluorescent yellow), IT186365 (yellow), IT137963 (whitish yellow), and IT180449 (greenish yellow). The total amount of carotenoid contents was in the order of orange color (104.64 mg/100 g), greenish yellow color (70.82 mg/100 g), fluorescent yellow color (32.41 mg/100 g), yellow color (8.73 mg/100 g), and whitish yellow color (4.73 mg/ 100 g). Both lutein and β-carotene were the predominant pigments of carotenoids, and lycopene was only separated and identified in the orange color flesh. According to the results, colorimetric analysis can aid breeders interested in increasing carotenoid content in squash, which could be accurately measured using a simple, reliable, and cost- and labor-efficient method for the evaluation of carotenoid pigments.