A study was conducted to investigate the effect of growing location in the physico-chemical properties of gum arabic obtained from Acacia senegal. Authentic representative gum exudates samples were collected from Abederafi, Yabello, Awash, and Filtu areas of Ethiopia. The experiment was conducted to determine Moisture content (MC), Ash content (Ash), pH, Relative Viscosity (RV), Nitrogen (N), Crude Protein (CP), and Mineral contents. Laboratory analyses were employed to determine the physico-chemical characteristics. The results were analyzed by using SAS Statistical Software. The analysis indicated that very high significant differences (p < 0.001) were seen on Ash, pH, RV, iron, calcium, and sodium contents of the gum-arabic samples between the study sites. Furthermore, significant differences (p < 0.05) were indicated for N, CP, Cu, and K contents. The results also showed MC, Mn, and Mg contents have no significant difference among growing locations. According to our results, the percentage moisture content of the gum-arabic samples were between 12.64% and 13.41%, while Ash percentage were between 0.32% and 16.98%. The mean pH values of the gum samples, obtained by employing different concentrations of aqueous solutions also ranged between 4.4 and 4.97. In addition, the relative viscosity of the gum samples studied falls between 0.9 to 4.2 cP, while Nitrogen and crude protein contents of the gum-arabic samples from the study areas were from 0.313% to 0.513% and 1.943% to 3.21%, respectively. The major mineral element compositions of the arabic gum samples in descending order were: Mg, Ca, Na, Cu, Fe, Mn, and K. As compared to international standards and prior studies on gum arabic, the gum-arabic samples from the study sites have comparable physico-chemical characteristics.