In addition to being widely used as a natural coloring material, β-carotene is an isoprenoid that acts as a pro-vitamin and antioxidant. It is a fat soluble molecule and hence soluble in organic solvents such as acetone, petroleum ether and methanol. The objective of the study was to determine the best extraction conditions for obtaining β-carotene from fresh pumpkin pulp using 3 different organic solvents (acetone, methanol and petroleum ether) and their combinations. The absorbance reading were measured in 454 nm in a Spectrophotometer (Jenway Model 6705 UV-Vis). Extraction solvents were pure acetone, methanol and petroleum ether and combinations were acetone:methanol (2:1) (v/v), methanol:ether (2:1), ether:acetone (2:1), all samples were analyzed in triplicate. Statistical analysis was performed for the Tukey test using the Software Assistat 7.7pt program. The results for β-carotene extraction were 148.72 μg/g for pure acetone; 19.8 μg/g for petroleum ether, 54.92 μg/g for pure methanol; 86.05 μg/g for petroleum ether:acetone (2:1); 94.85 μg/g for methanol: petroleum ether (2:1); 122.36 μg/g for acetone:methanol (2:1). Statistical analysis showed there was no significant difference (p<0,05) in the extraction performance between pure acetone and the acetone:methanol system (2:1). Based on the results, the best extractive solvent was pure acetone followed by acetone:methanol system (2:1) and the lowest extraction of β-carotene was obtained by pure petroleum ether. This fact can be explained by the solvent degree of affinity with the isoprenoid structure of β-carotene, among other factors such as solvent penetrability in the sample, incomplete extraction, losses by instability during extraction. In conclusion pure acetone showed to be the best solvent among all other extractive systems and this may be related to its high degree of affinity to the pumpkin pulp matrix.