AbstractThis paper proposes a method to determine bioethanol concentration that uses a pycnometer verified with a high performance liquid chromatography (HPLC) technique; it is a simple tool to determine the density of liquids for getting information about the ethanol concentration. The results showed that the sugar concentration affected the bioethanol concentration. A lower initial sugar concentration of 26.5 g/L generated higher yield of 45.3% sugar to bioethanol and a fractional or relative yield of 88.74%. Significance tests were used to compare the two experimental means, revealing that the pycnometer method and HPLC provide the same bioethanol concentration with joint variances of 2.269, 0.242, and 0.112 for 3 different tests with initial sugar concentrations of 26.486 g/L, 49.043 g/L, and 68.535 g/L, respectively. This study established and developed a methodology to determine bioethanol concentration from coffee mucilage by the proposed method.