Tej is an Ethiopian traditional alcoholic beverage with significant social and economic importance. Due to the spontaneous fermentation process of Tej, several issues such as safety, quality, and physicochemical properties of the final products is rquired to be assessed. Thus, this study was aimed to assess the microbial quality, physicochemical, and proximate properties of Tej associated with different maturity time. The microbial, physicochemical and proximate analyses were carried out by standard protocol. Lactic acid bacteria (6.30 log CFU/mL) and yeast (6.22 log CFU/mL) were the dominat microorganisms of all Tej samples at different maturity time, with significant differences (p = 0.001) in mean microbial count among samples. The mean pH, titratable acidity and ethanol content of Tej samples were 3.51, 0.79 and 11.04% (v/v), respectively. There were significant differences (p = 0.001) among the mean pH and titratable acidity values. The mean proximate compositions (%) of Tej samples were as follows: moisture (91.88), ash (0.65), protein (1.38), fat (0.47) and carbohydrate (3.91). Statistically significant differences (p = 0.001) were observed in proximate compositions of Tej samples from different maturity time. Generally, Tej maturity time has a great impact on the improvement of nutrient composition and the increment of the acidic contents which in turn suppress the growth of unwanted microorganisms. Further evaluation of the biological, and chemical safety and development of yeast-LAB starter culture are strongly recommended to improve Tej fermentation in Ethiopia.